Naphthalene ureas as glucose uptake enhancers

ABSTRACT

Compounds of formula I are useful for treating conditions associated with hyperglycemia, especially Type II diabetes. These compounds are useful in stimulating the kinase activity of the insulin receptor, activating the insulin receptor, and stimulating the uptake of glucose. Pharmaceutical compositions comprising the antidiabetic compounds are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority under 35 USC 119(e) of ProvisionalApplication No. 60/136,128, filed May 26, 1999.

BACKGROUND OF THE INVENTION

(a) Field of the Invention

The invention relates to a means to enhance insulin-dependent glucoseuptake. Specifically, the invention concerns compounds that activate theinsulin receptor kinase leading to increased glucose uptake. Theinvention also concerns methods for treating hyperglycemia in humans,and especially methods for treating Type II diabetes.

(b) Description of Related Art

Among the many functions performed by peptide and protein hormones inmetabolism is the ability to interact with receptors with highspecificity. The insulin receptor is present on virtually all cells andat high concentrations on the cells for the liver, skeletal muscles, andadipose tissue. Stimulation of the insulin receptor with insulin is anessential element in carbohydrate metabolism and storage.

Diabetics either lack sufficient endogenous secretion of the insulinhormone (Type I) or have an insulin receptor-mediated signaling pathwaythat is resistant to endogenous or exogenous insulin (Type II, ornon-insulin-dependent diabetes mellitus (NIDDM)). Type II diabetes isthe most common form of diabetes, affecting about 5% of individuals inthe industrialized nations. In Type II diabetics, majorinsulin-responsive tissues such as liver, skeletal muscle and fatexhibit the insulin resistance (Haring and Mehnert, Diabetologia36:176-182 (1993); Haring et al., Diabetologia, 37 Suppl. 2:S149-54(1994)). The resistance to insulin in Type II diabetes is complex andlikely multifactorial but appears to be caused by an impaired signalfrom the insulin receptor to the glucose transport system and toglycogen synthase. Impairment of the insulin receptor kinase has beenimplicated in the pathogenesis of this signaling defect. Insulinresistance is also found in many non-diabetic individuals, and may be anunderlying etiologic factor in the development of the disease (Reaven,Diabetes, 37:1595-1607 (1988)).

Considerable information is known concerning the insulin receptor itselfThe receptor consists of four separate subunits consisting of twoidentical a and two identical β chains. The β subunits contain atyrosine kinase activity and the ATP binding sites. The insulin receptoris activated by autophosphorylation of key tyrosine residues in itscytoplasmic tyrosine kinase domain. This autophosphorylation is requiredfor subsequent activity of the insulin receptor. The autophosphorylationstabilizes the activated receptor kinase resulting in a phosphorylationcascade involving intracellular signaling proteins.

At present there are limited pharmacologic approaches to treatment ofType II diabetes. Insulin is currently used as a treatment, but isdisadvantageous because insulin must be injected. Although severalpeptide analogs of insulin have been described, none with a molecularweight below about 5000 daltons retains activity. Some peptides whichinteract with sites on the β-subunit of the insulin receptor have shownenhancement of the activity of insulin on its receptor (Kole et al., J.Biol. Chem., 271:31619-31626 (1996); Kasuya et al., Biochem. Biophys.Res. Commun., 200:777-83 (1994)). Kohanski and others have reported on avariety of polycationic species that generate a basal effect, but dolittle to enhance insulin action (Kohanski, J. Biol. Chem., 264:20984-91(1989); Xu et al., Biochemistry 30:11811-19 (1991). These peptidesapparently act on the cytoplasmic kinase domain of the insulin receptor.

In addition, certain non-peptide components have been found to enhancethe agonist properties of peptide hormones, but none appear to actdirectly on the insulin receptor kinase. For instance, the ability ofthiazolidinediones, such as pioglitazone, to enhance adipocytedifferentiation has been described (Kletzien, et al., Mol. Pharmacol.,41:393 (1992)). These thiazolidinediones represent a class of potentialanti-diabetic compounds that enhance the response of target tissues toinsulin (Kobayashi, Diabetes, 41:476 (1992)). The thiazolidinedionesswitch on peroxisome proliferator-activated receptor γ (PPARγ), thenuclear transcription factor involved in adipocyte differentiation(Kliewer et al., J. Biol. Chem., 270:12953 (1995)) and do not have adirect effect on the insulin receptor kinase. Other anti-diabetic agentscurrently in use include both insulin secretagogues (such as thesulfonylureas) and biguanides (such as metfornin) that inhibit hepaticglucose output. To date, non-peptide substances which can mimic theactivating effect of insulin on the insulin receptor have eludeddiscovery.

Bisnaphthalene ureas are known to the literature. They are heavilydescribed as polysulfonic acid derivatives of suramin and as azo dyes. Avariety of these polyanionic sulfonic acid derivatives have beenestablished as potential therapeutics for a variety of diseaseindications. Suramin, described in 1917, is a polysulfonic acid that hasbeen extensively researched (Dressel, J. Chem. Ed., 38:585 (1961);Dressel, J. Chem. Ed., 39:320 (1962)). It has therapeutic uses as ananthehnintic and antiprotozoal. More recently, it has been described asan inhibitor to reverse transcriptase in certain avian and murineretroviruses (De Clercq, Cancer Letter, 8:9 (1979); Mitsuya et al.,Science, 226:172 (1984); Gagliardi et al., Cancer Chemother. Pharmacol.,41:117 (1988); Doukas et al., Cancer Res. 55:5161 (1995); Mohan et al.,Antiviral Chem., 2:215 (1991)). Large numbers of compounds relating tosuramin exist. Most of the suramin analogs which have been reported havemultiple sulfonic acid functionality on each aryl ring. Recent studiesindicate that polyanionic surarnin analogs have anti-angiogenic,antiproliferative activity, and anti-viral activity (Gagliardi et al.,Cancer Chemother. Pharmacol., 41:117 (1988); Doukas et al., Cancer Res.,55:5161 (1995); Mohan et al., Antiviral Chem., 2:215 (1991)). A numberof bisnaphthylsulfonic acids have been described in the patentliterature as complement inhibitors (U.S. Pat. Nos. 4,132,730,4,129,591, 4,120,891, 4,102,917, 4,051,176). Additionally, there are anumber of azo dye patents (DE 19521589, U.S. Pat. No. 3,716,368, DE2216592, FR 1578556) which disclose polysulfonated naphthalene azocompounds. Bisnaphthalene urea 2-sulfonamide 3-azo compounds have beensolely reported as a recording liquid (JP 58191772). However, none ofthe suramin analogs or azo dyes have been suggested to be useful in thetreatment of hyperglycemia or diabetes.

SUMMARY OF THE INVENTION

This invention is directed to bisnaphthalene ureas which enhance glucoseuptake in mammals, to pharmaceutical compositions thereof, and tomethods for enhancing glucose uptake using these compounds.

In a first embodiment, this invention provides compounds of formula I:

where

R¹ and R² are substituents on the A rings and are, independently,—SO₂NR⁷ ₂, —C(O)NR⁷ ₂, NR⁷SO₂R⁷, —NR⁷C(O)R⁷, —SO₂OR⁷, —C(O)OR⁷, —OSO₂R⁷,or —OC(O)R⁷,

R³ and R⁴ are, independently, hydrogen or lower alkyl, or R³ and R⁴together are —(CH₂)₂—, —(CH₂)₃—, or —(CH₂)₄—, or R³ or R⁴ may be anelectron pair,

R⁵ and R⁶ are, independently, hydrogen, alkyl, substituted alkyl, cyano,halo, nitro, —SR⁸, —C(O)R⁸, —SO₂OR⁸, —OSO₂R⁸, —SO₂NR⁸ ₂, —NR⁸SO₂R⁸,—OC(O)R⁸, —C(O)OR⁸, —C(O)NR⁸ ₂, —NR⁸C(O)R⁸, —OR⁸, or —NR⁸ ₂,

each R⁷ and R⁸ is, independently, hydrogen, alkyl, substituted alkyl,aryl, substituted aryl, aryl(lower)alkyl, substituted aryl(lower)alkyl,heteroaryl(lower)alkyl, substituted heteroaryl(lower)alkyl,heterocyclyl, substituted heterocyclyl, heteroaryl, or substitutedheteroaryl,

each Y is, independently, a non-interfering substituent which is notlinked to the naphthalene ring via an azo or amide linkage,

each x is, independently, 0, 1 or 2, and

linker connects a carbon designated as c to a carbon designated as d,and is

 where K is O, S, or NR^(*), and R^(*) is H, cyano, or lower alkyl; or

 where R^(*) is H or lower alkyl; or

 where R^(*) is H, cyano, or lower alkyl;

 where R^(*) is cyano or lower alkyl;

and, where, if R¹ and R² are both —SO₂OH, then:

(i) no Y is —SO₂OH;

(ii) neither R⁵ nor R⁶ is —SO₂OR⁸ or —OSO₂R⁸; and

(iii) R⁵ and R⁶ are not both selected from the group consisting ofhydroxy and hydrogen unless at least one (Y)_(x) is (Y′)_(x′) where x′is 1 or 2 and Y′is a halo, and the pharmaceutically acceptable saltsthereof, as single stereoisomers or mixtures of stereoisomers. Thesecompounds are useful as glucose uptake agonists and in the treatment ofhyperglycemia and diabetes.

In a second embodiment, this invention provides pharmaceuticalcompositions comprising (a) a pharmaceutically acceptable carrier and(b) as an active ingredient, a compound of the first embodiment. Thesecompositions are useful for stimulating the uptake of glucose into cellsin a mammal or for treating a mammalian disease state selected from thegroup consisting of hyperglycemia, type I diabetes, and type IIdiabetes.

In a third embodiment, this invention provides a method of stimulatingthe kinase activity of the insulin receptor, comprising contacting theinsulin receptor, or the kinase portion thereof, with a compound of thefirst embodiment, in an amount sufficient to stimulate the kinaseactivity of the insulin receptor.

In a fourth embodiment, this invention provides a method of activatingthe insulin receptor is provided, comprising contacting the insulinreceptor, or the kinase portion thereof, with a compound of the firstembodiment, in an amount sufficient to effect activation of the insulinreceptor.

In a fifth embodiment, this invention provides a method for stimulatingthe uptake of glucose into cells which display the insulin receptor,involving contacting the cells, optionally in the presence of insulin,with a compound of the first embodiment, in an amount sufficient tostimulate the uptake of glucose into the cells. The uptake of glucoseinto cells in a mammal may be effected by administering the compound ofthe invention to the mammal.

In other embodiments, the invention provides methods of treatinghyperglycemia, type I diabetes, or type II diabetes in a mammal, such asa human by administering a therapeutically effective amount of acompound of the first embodiment or a composition containing saidcompound.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the phosphorylation of IRS-1 and the insulin receptor withCompound 15 with and without insulin.

FIG. 2 shows the increase in the phosphorylation of the insulin receptorwhen treated with Compounds 13 and 15 at various concentrations.

FIG. 3 shows the increase in glucose uptake of cells when treated withCompounds 13 or 15 in the presence or absence of insulin.

FIG. 4 shows the glucose transport effect of Compound 13 or 15 in thepresence or absence of wortmannin.

FIG. 5 shows the glucose transport effect of Compound 15 in the presenceor absence of Wortmannin and Cytochalasin B.

FIG. 6 shows GLUT4 immunofluorescence of cells when treated withCompound 15.

FIG. 7 shows the effect of Compound 15 against the insulin and EGFreceptor.

FIG. 8 shows the blood glucose lowering effect of Compound 15 withinsulin in a db/db mouse.

FIG. 9 shows the blood glucose lowering effect of Compound 15 only in adb/db mouse.

FIG. 10 shows the effect of Compound 15 on certain blood components inan ob/ob mouse.

FIG. 11 shows the effect of Compound 15 on blood glucose levels in aSTZ/BFD rat.

FIG. 12 shows the amount of insulin receptor phosphorylation found inmuscle tissue after oral administration of Compound 15.

FIG. 13 shows the effect of Compound 15 after 3 single daily doses inthe db/db mouse.

FIG. 14 shows the effect of Compound 15 after 3 single daily doses inthe STZ/HFD rat.

DETAILED DESCRIPTION OF THE INVENTION

(a) Definitions and General Parameters

“Alkyl”, as in “alkyl”, or “alkyloxy”, means C₁-C₂₀ monovalenthydrocarbyl moiety which may be linear, branched, or cyclic. “Loweralkyl”, as in “lower alkyl” “halo-lower alkyl”, “aryl(lower)alkyl”, or“heteroaryl(lower)alkyl”, means a C₁-C₆ alkyl. The term “lower akyl”includes such moieties as methyl, ethyl, isopropyl, propyl, butyl,isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, cyclopentyl,cyclopropylmethyl, or cyclohexyl. C₁-C₄ lower alkyls are preferred.“Lower alkyl” as used herein includes moieties with one olefinic bond,such as allyl.

A “substituted alkyl” or “substituted lower alkyl” is an alkyl or loweralkyl, respectively, which is typically mono-, di-, or trisubstitutedwith a moiety such as aryl, R′-substituted aryl, heteroaryl, niitro,cyano, halo, —OR, —SR, —COR, —OC(O)R, —C(O)OR, —NR₂, —SO₂OR, —OSO₂R,—SO₂NR₂, —NRSO₂R, —CONR₂, or —NRCOR, where each R is, independently,hydrogen, lower alkyl, R′-substituted lower alkyl, aryl, R′-substitutedaryl, heteroaryl, heteroaryl(lower)alkyl, R′-substitutedaryl(lower)alkyl, or aryl(lower)alkyl and each R′ is, independently,hydroxy, halo, lower alkyloxy, cyano, thio, nitro, lower alkyl,halo-lower alkyl, or amino. Substituted alkyls or substituted loweralkyls which are substituted with one to three of the substituentsselected from the group consisting of cyano, halo, lower alkyloxy, thio,nitro, amino, or hydroxy are particularly preferred.

The term “halo-lower alkyl” means a lower alkyl substituted with one tothree halo groups, and is further exemplified by such radicals as —CF₃,—CH₂CF₃ and —CH₂CCl₃.

“Aryl”, as in “aryl”, “aryloxy”, and “aryl(lower)alkyl”, means a radicalderived from an aromatic hydrocarbon containing 6 to 20 ring carbonatoms, having a single ring (e.g., phenyl), or two or more condensedrings, preferably 2 to 3 condensed rings (e.g., naphthyl), or two ormore aromatic rings, preferably 2 to 3 aromatic rings, which are linkedby a single bond (e.g., biphenyl). The aryl is preferably C₆-C₁₆ andeven more preferably, C₆ to C₁₄.

A “substituted aryl” is an aryl radical which is mono-, di-, ortrisubstituted, independently, with a moiety such as a hydroxy,triazolyl, tetrazolyl, hydroxyisoxazolyl, phosphonic acid or phosphonateresidue, alkyl, R′-substituted alkyl, halo, trifluoromethyl, cyano,nitro, —SR, —OR, —COR, —OCOR, —SO₂OR, —OSO₂R, —SO₂NR₂, —NRSO₂R, —COOR,—NR₂, —CONR₂, or —NRCOR, where each R is, independently, hydrogen, loweralkyl, R′-substituted lower alkyl, aryl, R′-substituted aryl,heteroaryl, heteroaryl(lower)alkyl, aryl(lower)alkyl, or R′-substitutedaryl(lower)alkyl and each R′ is, independently hydroxy, halo, loweralkyloxy, cyano, thio, nitro, lower alkyl, halo-lower alkyl, amino, or—COOR, where R is as defined above. Especially preferred substituents ona substituted aryl are lower alkyl, halo-lower alkyl, halo, cyano, thio,nitro, amino, lower alkyloxy, or hydroxy. The radicals —SO₂OR, —SO₂NR₂,—COOR, and —CONR₂, where R is hydrogen or lower alkyl, are alsoespecially preferred substituents of substituted aryls on the compoundsof the present invention.

“Heteroaryl”, as in “heteroaryl” and “heteroaryl(lower)alkyl”, means aradical derived from an aromatic hydrocarbon containing 5 to 14 ringatoms, 1 to 5 of which are hetero atoms chosen, independently, from N,O, or S, and includes monocyclic, condensed heterocyclic, and condensedcarbocyclic and heterocyclic aromatic rings (e.g., thienyl, fliryl,pyrrolyl, pyrimidinyl, isoxazolyl, oxazolyl, triazolyl, tetrazolyl,indolyl, isobenzofuranyl, purinyl, isoquinolyl, pteridinyl, imidazolyl,pyridyl, pyrazolyl, pyrazinyl, quinolyl, etc.).

A “substituted heteroaryl” may have from one to three substituents suchas an alkyl, R′-substituted alkyl, halo, cyano, nitro, —SR, —OR, —COR,—OOCR, —SO₂OR, —OSO₂R, —SO₂NR₂, —NRSO₂R, —COOR, —NR₂, —CONR₂, or —NRCOR,where each R is independently hydrogen, lower alkyl, R′-substitutedlower alkyl, aryl, R′-substituted aryl, heteroaryl,heteroaryl(lower)alkyl, aryl(lower)alkyl, or R′-substitutedaryl(lower)alkyl and each R′ is, independently, hydroxy, halo, loweralkyloxy, cyano, thio, nitro, lower alkyl, halo-lower alkyl, or amino.In addition, any two adjacent substituents on the heteroaryl mayoptionally together form a lower alkylenedioxy. Particularly preferredsubstituents on the substituted heteroaryl include hydroxy, halo, loweralkyloxy, cyano, thio, nitro, lower alkyl, halo-lower alkyl, halo-loweralkyl, or amino.

“Heterocyclyl” means a radical derived from an aliphatic, cyclichydrocarbon containing 5 to 14 ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S. Monocyclic rings (e.g.,tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, etc.) are preferred.

A “substituted heterocyclyl” may have from one to three substituents,preferably substituents like an alkyl, R′-substituted alkyl, halo,cyano, nitro, —SR, —OR, —COR, —OOCR, —SO₂OR, —OSO₂R, —SO₂NR₂, —NRSO₂R,—COOR, —NR₂, —CONR₂, or —NRCOR, where each R is, independently,hydrogen, loweralkyl, R′-substituted alkyl, aryl, R′-substituted aryl,heteroaryl, heteroaryl(lower)alkyl, aryl(lower)alkyl, or R′-substitutedaryl(lower)alkyl and each R′ is, independently hydroxy, halo, loweralkyloxy, cyano, thio, nitro, lower alkyl, halo-lower alkyl, or aimino.Preferred substituents on a substituted heterocyclyl include loweralkyl, halo-lower alkyl, halo, cyano, thio, amino, lower alkyloxy, orhydroxy.

“Aryl(lower)alkyl” means a lower alkyl radical which is substituted withan aryl, as previously defined. A “substituted aryl(lower)alkyl” meansan aryl(lower)alkyl radical having one to three substituents on the arylportion or the alkyl portion of the radical, or both.

“Heteroaryl(lower)alkyl” means a lower alkyl radical which issubstituted with a heteroaryl, as previously defined. A “substitutedheteroaryl(lower)aryl” means a heteroaryl(lower)alkyl radical having oneto three substituents on the heteroaryl portion or the alkyl portion ofthe radical, or both.

A “lower alkyloxy” means an —OR radical, where R is a lower alkyl.

“Halo” means bromo, fluoro, or chloro.

A “non-interfering substituent” means a substituent which, when presenton a given compound, does not substantially decrease or otherwiseinhibit a particular, desired bioactivity of the compound, such as theability of the compound to stimulate the kinase activity of the insulinreceptor, to activate the insulin receptor, or to stimulate the uptakeof glucose into cells displaying the insulin receptor. The presence ofthe non-interfering substituent should not detrimentally affect thebioactivity of the compound by more than about 30%. Preferably, thenon-interfering substituent decreases the bioactivity of the compound byless than about 10%. Most preferably, the non-interfering substituentdoes not decrease the bioactivity of the compound to any detectabledegree. However, the effect of the presence of the non-interferingsubstituent on the compound need not be neutral. For instance, thenon-interfering substituent may optionally increase a particularbioactivity of the compound. Suitable non-interfering substituentsinclude, but are not limited to, alkyl, substituted alkyl, cyano, halo,nitro, —SR, —OR, and —NR₂, where each R is, independently, hydrogen,lower alkyl, or substituted lower alkyl.

An “azo linkage” is the group —N═N—. A typical “amnide linkage” is thegroup

where R may be alkyl, aryl, or hydrogen.

A “pharmaceutically acceptable salt” may be any salt derived from aninorganic or organic acid or an inorganic or organic base. The term“pharmaceutically acceptable anion” refers to the anion of such acidaddition salts. The term “pharmaceutically acceptable cation” refers toa cation formed by addition of a base. The salt and/or the anion orcation are chosen not to be biologically or otherwise undesirable.

“Stereoisomers” are compounds that have the same sequence of covalentbonds and differ in the relative disposition of their atoms in space.

“Inner salts”0 or “zwitterions” can be formed by transferring a protonfrom the carboxyl group onto the lone pair of electrons of the nitrogenatom in the amino group.

“Therapeutically effective amount” means that amount which, whenadministered to a mammal for treating a disease, is sufficient to effectsuch treatment for the disease.

“Treating” or “treatment” of a disease in a mammal includes:

(1) preventing the disease from occurring in a mammal which may bepredisposed to the disease but does not yet experience or displaysymptoms of the disease,

(2) inhibiting the disease, i.e., arresting its development, or

(3) relieving the disease, i.e., causing regression of the disease.

The “kinase portion thereof”, with respect to the insulin receptor,means the cytoplasmic tyrosine kinase domain of the insulin receptor.

(b) Nomenclature

The compounds of formula I are numbered and named as described belowwith reference to formula Ia.

In the compound of formula Ia shown, the substituent R¹ is in the7-position of the naphthalene ring, and R² is in the 2-position of thenaphthalene ring when the numbering of the ring atoms is as shown. Forexample, if R¹ is SO₂OH, R² is SO₂OH, R³ is H, R⁴ is H, R⁵ is H, R⁶ isH, and K is O, and the aminocarbonylamino linker is attached to C2 ofthe naphthalene ring with the R⁵ substituent and is attached to C7 ofthe naphthalene ring with the R⁶ substituent on it, the compound is7-{[(7-sulfo-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic acid.

(c) Compounds and Pharmaceutical Compositions Thereof

The compounds of the invention comprise compounds of formula I:

where

R¹ and R² are substituents on the A rings and are, independently,—SO₂NR⁷ ₂, C(O)NR⁷ ₂, —NR⁷SO₂R⁷, —NR⁷C(O)R⁷, —SO₂OR⁷, —C(O)OR⁷, —OSO₂R⁷,or —OC(O)R⁷,

R³ and R⁴ are, independently, hydrogen or lower alkyl, or R³ and R⁴together are —(CH₂)₂—, —(CH₂)₃—, or —(CH₂)₄—, or R³ or R⁴ may be anelectron pair, p1 R⁵ and R⁶ are, independently, hydrogen, alkyl,substituted alkyl, cyano, halo, nitro, —SR⁸, —C(O)R⁸, —SO₂OR⁸, —OSO₂R⁸,—SO₂NR⁸ ₂, —NR⁸SO₂R⁸, —OC(O)R⁸, —C(O)OR⁸, —C(O)NR⁸ ₂, —NR⁸C(O)R⁸, —OR⁸,or —NR⁸ ₂,

each R⁷ and R⁸ is, independently, hydrogen, alkyl, substituted alkyl,aryl, substituted aryl, aryl(lower)alkyl, substituted aryl(lower)alkyl,heteroaryl(lower)alkyl, substituted heteroaryl(lower)alkyl,heterocyclyl, substituted heterocyclyl, heteroaryl, or substitutedheteroaryl,

each Y is, independently, a non-interfering substituent which is notlinked to the naphthalene ring via an azo or amide linkage,

each x is, independently, 0, 1 or 2, and

the linker connects a carbon designated as c to c carbon designated asd, and is

 where K is O, S, or NR^(*), and R^(*) is H, cyano, or lower alkyl; or

 where R^(*) is H or lower alkyl; or

 where R^(*) is H, cyano, or lower alkyl;

 where R^(*) is cyano or lower alkyl;

and, where, if R¹ and R² are both —SO₂OH, then:

(i) no Y is —SO₂OH;

(ii) neither R⁵ nor R⁶ is —SO₂OR⁸ or OSO₂R⁸; and

(iii) R⁵ and R⁶ are not both selected from the group consisting ofhydroxy and hydrogen unless at least one (Y)_(x) is (Y′)_(x′) where x′is 1 or 2 and Y′ is a halo,

and the pharmaceutically acceptable salts thereof,

as single stereoisomers or mixtures of stereoisomers.

Preferred compounds of formula I include compounds of formula II:

where the substituents and linker are as defined above,

and pharmaceutically acceptable salts thereof, as single stereoisomersor mixtures of stereoisomers.

Preferably, the compounds of formula II include compounds of formula IIa

where the substituents and linker are as defined above,

and pharmaceutically acceptable salts thereof, as single stereoisomersor mixtures of stereoisomers.

Preferably, each non-interfering substituent Y is alkyl, substitutedalkyl, cyano, halo, nitro, —SR⁹, —OR⁹, or —NR⁹ ₂, where each R⁹ is,independently, hydrogen, lower alkyl, or substituted lower alkyl. Mostpreferably, each Y is lower alkyl, halo-lower alkyl, lower alkyloxy,cyano, halo, thio, nitro, amino, or hydroxy.

In the compounds of formula I, Ia, II, and IIa, each x is preferablyzero or one. In particularly preferred embodiments, each x is zero.

In one preferred embodiment of the compounds of the invention, R¹ and R²are, independently, —SO₂OR¹⁰, —C(O)OR¹⁰, —SO₂NR¹¹R¹⁰, —C(O)NR¹¹R¹⁰,—OSO₂R¹⁰, —OC(O)R¹⁰, —NR¹¹SO₂R¹⁰, or —NR¹¹C(O)R^(10; each R) ¹¹ is,independently, hydrogen or lower alkyl; and each R¹⁰ is, independently,alkyl, substituted alkyl, aryl, substituted aryl, aryl(lower)alkyl,substituted aryl(lower)alkyl, heteroaryl(lower)alkyl, substitutedheteroaryl(lower)alkyl, heterocyclyl, substituted heterocyclyl,heteroaryl, or substituted heteroaryl. R¹ and R² are preferably,independently, —SO₂NR¹¹R¹⁰, —C(O)NR¹¹R¹⁰, —NR¹¹SO₂R¹⁰ , or —NR¹¹C(O)R¹⁰.In a further preferred embodiment, R¹¹ is hydrogen. For instance, inparticularly preferred compounds, R¹ and R² are, independently,—SO₂NHR¹⁰ or —NHSO₂R¹⁰.

In an alternative preferred embodiment, R¹ is —SO₂OR¹⁰, —C(O)OR¹⁰,—SO₂NR¹¹R¹⁰, —C(O)NR¹¹R¹⁰, —OSO₂R¹⁰, —OC(O)R¹⁰, —NR¹¹SO₂R¹⁰, or—NR¹¹C(O)R¹⁰, and R² is —SO₂OR¹¹, or —C(O)OR¹¹, where R¹⁰ and R¹¹ are aspreviously defined in the preceding paragraph. R¹ is, preferably,—SO₂NR¹¹R¹⁰, —C(O)NR¹¹R¹⁰, —NR¹¹SO₂R¹⁰, or —NR¹¹C(O)R¹⁰. In a furtherpreferred embodiment, R¹¹ is hydrogen. For instance, in particularlypreferred compounds, R¹ is —SO₂NHR¹⁰ or —NHSO₂R¹⁰. In one preferredembodiment, R² is —C(O)NR¹¹ ₂ or —C(O)OR¹¹. In an alternative preferredembodiment, R² is —SO₂NR¹¹ ₂ or —SO₂OR¹¹, such as —SO₂OH.

In one preferred embodiment of the invention, each R¹⁰ is,independently, a substituted alkyl, substituted aryl, substitutedaryl(lower)alkyl, substituted heteroaryl(lower)alkyl, substitutedheterocyclyl, or substituted heteroaryl; at least one of thesubstituents on R¹⁰ is R¹²; each R² is, independently, —SO₂OR¹³,—C(O)OR¹³, —SO₂NR¹³ ₂, —C(O)NR¹³ ₂, hydroxy, triazolyl, tetrazolyl,hydroxyisoxazolyl, a phosphonic acid residue, or a phosphonate residue;and each R¹³ is, independently, hydrogen or lower alkyl. R¹⁰ in thisembodiment is preferably a substituted aryl or substituted heteroaryl.It is particularly preferred that R¹⁰ be a substituted phenyl. Inpreferred compounds of the invention, each R¹² is, independently,—C(O)OR¹³, —C(O)NR¹³ ₂, —SO₂OR¹³, hydroxy, triazolyl, tetrazolyl,hydroxyisoxazolyl, a phosphonic acid residue, or a phosphonate residue.In particular, it is preferred that R¹² be —C(O)OR¹³, —SO₂OR¹³, hydroxy,triazolyl, tetrazolyl, hydroxyisoxazolyl, a phosphonic acid residue, ora phosphonate residue. For instance, each R¹² may be —C(O)OH, —SO₂OR¹³,or —C(O)OCH₃. In a alternatively preferred compounds, each R¹² is,independently, —SO₂OR¹³ or —SO₂NR¹³ ₂. It is particularly preferred thatR¹² be —SO₂OR¹³. In especially preferred compounds, R¹² is —SO₂OH. It ispreferred that, when R¹² is —C(O)OR¹³ or —SO₂OR¹³, R¹² shall be adjacenton the aryl, heteroaryl, or heterocyclyl ring to a substituent such aschloro or hydroxy.

In an alternative preferred embodiment of the invention, each R¹⁰ is,independently, an aryl, heteroaryl, aryl(lower)alkyl, orheteroaryl(lower)alkyl. In this embodiment, R¹⁰ is preferably phenyl,pyridyl, pyrazinyl, or pyrimidinyl.

In still other preferred compounds of the invention, each R¹ and R² are,independently, —SO₂NR⁷ ₂, —C(O)NR⁷ ₂, —SO₂OR⁷, or —C(O)OR⁷; and each R⁷is, independently, hydrogen or lower alkyl. Preferably, R¹ and R² are,independently, —C(O)OR⁷ or —C(O)NR⁷ ₂. In other preferred compounds offormula I, however, R¹ and R² are, independently, —SO₂OR⁷ or —SO₂NR⁷ ₂.For instance, R¹ and R² may both be —SO₂OH. Preferably, if R¹ and R² areboth —SO₂OH then R⁵ and R⁶ are not both either hydroxy or hydrogen.

Preferably, R³ and R⁴ of the compounds of the invention are hydrogen.

Preferably, R⁵ and R⁶ are independently, hydrogen, alkyl, substitutedalkyl, cyano, halo, nitro, —OR⁸, ^(NR) ⁸ ₂, or —SR⁸, where each R⁸ is,independently, hydrogen, lower alkyl, substituted lower alkyl, aryl,substituted aryl, aryl(lower)alkyl, substituted aryl(lower)alkyl,heteroaryl, or heteroaryl(lower)alkyl. Most preferably, R⁵ and R⁶ are,independently, hydrogen, hydroxy, halo, cyano, lower alkyl, halo-loweralkyl, lower alkyloxy, nitro, amino, or thio. In many preferredcompounds of the invention, R⁵ and R⁶ are both hydrogen or hydroxy.

The compounds of formula I and formula II are preferably symmetrical.

Compounds of the invention comprising more than one preferredsubstituent are preferred. If one compound comprises more preferredsubstituents than a second compound, then the first compound ispreferred over the second. For instance, compounds of formula I whichhave preferred radicals for each substituent R¹ through R⁴ and R¹⁰through R¹² are preferred over those which have preferred radicals foronly the substituents R¹ through R³.

For example, preferred compounds of the invention include those offormula III:

where R⁵ and R⁶ are selected from the group consisting of hydrogen andhydroxy; and each R¹⁰ is, independently, substituted aryl or substitutedheteroaryl; at least one of the substituents on R¹⁰ is R¹²; each R¹² is,independently, —SO₂OR¹³, —C(O)OR¹³, —SO₂NR¹³ ₂, —C(O)NR¹³ ₂, triazolyl,tetrazolyl, isoxazolyl, a phosphonic acid residue, or a phosphonateresidue; and each R¹³ is, independently, hydrogen or lower alkyl, andpharmaceutically acceptable salts thereof as single stereoisomers ormixtures of stereoisomers.

Preferably, when R¹² is —CO(O)R¹³ or —SO₂OR¹³, R¹² is adjacent on thearyl, heteroaryl, or heterocyclyl ring to a further substituent, such aschloro or hydroxy.

Other examples of compounds of the present invention include those offormula IV:

where R⁵ and R⁶ are selected from the group consisting of hydrogen andhydroxy; R¹⁰ is substituted aryl or substituted heteroaryl; at least oneof the substituents on R¹⁰ is R¹²; each R¹² is, independently, —SO₂OR¹³,—C(O)OR¹³, —SO₂NR¹³ ₂, —C(O)NR¹³ ₂, triazolyl, tetrazolyl, isoxazol, aphosphonic acid residue, or a phosphonate residue; and each R¹³ is,independently, hydrogen or lower alkyl,

and pharmaceutically acceptable salts thereof as single stereoisomers ormixtures of stereoisomers. Preferably, when R¹² is —CO(O)R¹³ or—SO₂OR¹³, R¹² shall be adjacent on the aryl, heteroaryl, or heterocyclylring to a substituent such as chloro or hydroxy.

Preferred compounds of the present invention include, but are notlimited to, the following compounds:

4-hydroxy-7-{[(5-hydroxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid disodium salt;

3-{[(4-hydroxy-7-{[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]-carbonylamino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid;

2-[6-({[5-(carboxymethoxy)-7-sulfo(2-naphthyl)]amino}carbonylamino)-3-sulfonaphthyloxy]aceticacid;

3-bromo-7-{[(6-bromo-5-hydroxy-7-sulfo(2-naphthyl))amino]carbonylamino}-4-hydroxynaphthalene-2-sulfonicacid;

4-[(2-sulfophenyl)methoxy]-7-[({7-sulfo-5-[(2-sulfophenyl)methoxy](2-naphthyl)}amino)-carbonylamino]naphthalene-2-sulfonicacid;

4-hydroxy-7-{[(5-methoxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid;

4-methoxy-7-{[(5-methoxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid;

7-[({5-[(ethoxycarbonyl)methoxy]-7-sulfo(2-naphthyl)}amino)carbonylamino]-4-hydroxynaphthalene-2-sulfonicacid;

4-[(ethoxycarbonyl)methoxy]-7-[({5-[(ethoxycarbonyl)methoxy]-7-sulfo(2-naphthyl)}amino)-carbonylamino]naphthalene-2-sulfonicacid;

4-(3-sulfopropoxy)-7-({[7-sulfo-5-(3-sulfopropoxy)(2-naphthyl)]amino}carbonylamino)naphthalene-2-sulfonicacid;

4-hydroxy-7-({[7-sulfo-5-(3-sulfopropoxy)(2-naphthyl)]amino}carbonylamino)naphthalene-2-sulfonmcacid;

N-(5-hydroxy-7-sulfamoyl(2-naphthyl))[(5-hydroxy-7-sulfamoyl(2-naphthyl))amino]-carboxamide;

4-hydroxy-7-{[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]-carbonylamino}naphthalene-2-sulfonicacid;

methyl3-({[4-hydroxy-7-({[5-hydroxy-7-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl)-(2-naphthyl)]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate;

3-{[(7-{[(7-{[(3-carboxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))amino]-carbonylamino}-4-hydroxy-2-naphthyl)sulfonyl]amino}benzoicacid;

4-{[(7-{[(7-{[(4-carboxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))amino]-carbonylamino}-4-hydroxy-2-naphthyl)sulfonyl]amino}benzoicacid;

4-{[(4-hydroxy-7-{[N-(5-hydroxy-7-sulfo(2-naphthyl))carbamoyl]amino}-2-naphthyl)sulfonyl]-amino}benzoicacid;

methyl4-({[4-hydroxy-7-({N-[5-hydroxy-7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)-(2-naphthyl)]carbamoyl}amino)-2-naphthyl]sulfonyl}amino)benzoate;

4-hydroxy-7-({[5-hydroxy-7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)naphthalene-2-sulfonicacid;

7-{[(7-sulfo-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic acid;

7-{[(7-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonicacid;

3-{[(7-{[N-(7-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)-sulfonyl]amino}benzenesulfonicacid;

methyl3-({[7-({[7-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl)-2-naphthyl]amino}-carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate;

3-{[(7-{[N-(7-{[(3-carboxyphenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)-sulfonyl]amino}benzoicacid;

N-{7-[(phenylamino)sulfonyl](2-naphthyl)}({7-[(phenylamino)sulfonyl](2-naphthyl)}amino)carboxamide;

N-(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide;

N-{7-[(3-pyridylamino)sulfonyl](2-naphthyl)}({7-[(3-pyridylamino)sulfonyl]-2-naphthyl}amino)carboxamide;

N-{7-[(pyrazin-2-ylamino)sulfonyl](2-naphthyl)}({7-[(pyrazin-2-ylamino)sulfonyl]-2-naphthyl}amino)carboxamide;

N-{7-[(pyrimidin-2-ylamino)sulfonyl](2-naphthyl)}({7-[(pyrimidin-2-ylamino)sulfonyl]-2-naphthyl}amino)carboxamide;

4-methylphenyl3-[({7-[(N-{7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate;

4-methylphenyl4-[({7-[({7-[({4-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}amino)carbonylamino]-2-naphthyl}sulfonyl)amino]benzenesulfonate;

7-[({7-[(pyrimidin-2-ylamino)sulfonyl]-2-naphthyl}amino)carbonylamino]naphthalene-2-sulfonicacid;

4-{[(7-{[N-(7-{[(4-sulfophenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphtyl)-sulfonyl]amino}benzenesulfonicacid;

methyl4-({[7-({N-[7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)-2-naphthyl]carbamoyl}amino)-2-naphthyl]sulfonyl}amino)benzoate;

4-{[(7-{[N-(7-{[(4-carboxyphenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2naphthyl)-sulfonyl]amino}benzoicacid;

methyl(2S)-2-({[7-({N-[7-({[(1S)-2-(4-hydroxyphenyl)-1-(methoxycarbonyl)ethyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)-3-(4-hydroxyphenyl)propanoate;and

(2S)-2-({[7-({N-[7-({[(1S)-1-carboxy-2-(4-hydroxyphenyl)ethyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)-3-(4-hydroxyphenyl)propanoicacid;

and their pharmaceutically acceptable salts, as single stereoisomers ormixtures of stereoisomers.

Syntheses and descriptions of these compounds are outlined in Examples 1through 10 below.

Certain compounds of the invention may contain one or more chiralcenters. In such cases, all stereoisomers also fall within the scope ofthis invention. The invention compounds include the individuallyisolated stereoisomers as well as mixtures of such stereoisomers.

The compounds of the invention further comprise pharmaceuticallyacceptable salts of the compounds disclosed herein. Thesepharmaceutically acceptable salts are suitable for use in all methodsand pharmaceutical compositions of the present invention.

Pharmaceutically acceptable salts include salts which may be formed whenacidic protons present are capable of reacting with inorganic or organicbases. Typically the parent compound is treated with an excess of analkaline reagent, such as hydroxide, carbonate or alkoxide, containingan appropriate cation. Cations such as Na⁺, K⁺, Ca²⁺ and NH₄ ⁺ areexamples of cations present in pharmaceutically acceptable salts. TheNa⁺ salts are especially useful. Acceptable inorganic bases, therefore,include calcium hydroxide, potassium hydroxide, sodium carbonate andsodium hydroxide. Salts may also be prepared using organic bases, suchas ethanolamine, diethanolamine, triethanolamine, N-methylglucamine,ethanolamine, and tromethamine.

If a compound of the invention contains a basic group, an acid additionsalt may be prepared. Acid addition salts of the compounds are preparedin a standard manner in a suitable solvent from the parent compound andan excess of an acid, such as hydrochloric acid, hydrobromic acid,sulfuric acid (giving the sulfate and bisulfate salts), nitric acid,phosphoric acid and the like, and organic acids such as acetic acid,propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid,malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid,citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonicacid, ethanesulfonic acid, salicylic acid, p-toluenesulfonic acid,hexanoic acid, heptanoic acid, cyclopentanepropionic acid, lactic acid,o-(4-hydroxy-benzoyl)benzoic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,p-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,camphorsulfonic acid, 4-methyl-bicyclo[2.2.2.]oct-2-ene-1-carboxylicacid, glucoheptonic acid, gluconic acid,4,4′-methylenebis(3-hydroxy-2-naphthoic)acid, 3-phenylpropionic acid,trimethylacetic acid, t-butylacetic acid, laurylsulfuric acid,glucuronic acid, glutamic acid, 3-hydroxy-2-naphthoic acid, stearicacid, muconic acid and the like.

Certain of the compounds of the invention form inner salts orzwitterions.

The invention includes pharmaceutical compositions of all the compoundsof the present invention. These pharmaceutical compositions comprise (i)a compound of the invention as an active ingredient and (ii) apharmaceutically acceptable carrier.

Pharmaceutical compositions of the compounds of this invention, orderivatives thereof, may be formulated as solutions or lyophilizedpowders for parenteral administration. Powders may be reconstituted byaddition of a suitable diluent or other pharmaceutically acceptable,carrier prior to use. The liquid formulation is generally a buffered,isotonic, aqueous solution. Examples of suitable diluents are normalisotonic saline solution, 5% dextrose in water or buffered sodium orammonium acetate solution. Such formulations are especially suitable forparenteral administration, but may also be used for oral administration.It may be desirable to add excipients such as polyvinylpyrrolidinone,gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodiumchloride or sodium citrate. Alternatively, these compounds may beencapsulated, tableted or prepared in an emulsion or syrup for oraladministration. Pharmaceutically acceptable solid or liquid carriers maybe added to enhance or stabilize the composition, or to facilitatepreparation of the composition. Liquid carriers include syrup, peanutoil, olive oil, glycerin, saline, alcohols and water. Solid carriersinclude starch, lactose, calcium sulfate dihydrate, terra alba,magnesium stearate or stearic acid, talc, pectin, acacia, agar orgelatin. The carrier may also include a sustained release material suchas glyceryl monostearate or glyceryl distearate, alone or with a wax.The amount of solid carrier varies but, preferably, will be betweenabout 20 mg to about 1 g per dosage unit. The pharmaceuticalpreparations are made following the, conventional techniques of pharmacyinvolving milling, mixing, granulation, and compressing, when necessary,for tablet forms; or milling, mixing and filling for hard gelatincapsule forms. When a liquid carrier is use, the preparation will be inthe form of a syrup, elixir, emulsion or an aqueous or non-aqueoussuspension. Such a liquid formulation may be administered directly p.o.or filled into a soft gelatin capsule. On a percent by weight basis,typical pharmaceutical compositions may contain from 0.1 to 95% ofactive ingredient, more preferably 1 to 80%.

Some specific examples of suitable pharmaceutical compositions aredescribed in Example 25 below.

Typically, a pharmaceutical composition of the present invention wouldbe packaged in a container with a label indicating use of thepharmaceutical composition in the treatment of hyperglycemia, type Idiabetes, and type II diabetes, or a combination of any of these diseaseconditions. Compounds can be prophylactically administered prior to ameal to control excessive elevated glucose in type II diabetics for aperiod of time following the meal. Alternatively, the compounds can beadministered to a diabetic to normalize excessively elevated glucoselevels as measured by glucose monitoring device. Finally, it is knownthat the very small amount of residual circulating insulin found in typeI diabetics is used by the body to prevent severe lipolysis and theresulting keto-acidosis. These compounds can be used to prophylacticallyagainst ketoacidosis in type I patient. Administration of thesecompounds could provide relief from ketoacidosis via insulin receptoractivation, not for control of blood sugar, but for inhibition of severelipolysis in type I diabetics who do not have ready access to insulin.

(c) Methods of Use of the Compounds of the Present Invention

Compounds of the present invention have been found to stimulateautophosphorylation of the insulin receptor (Example 11 and 12, below).In addition, these compounds have been shown to enhance insulin'sability to effect the transport of glucose into cultured fibroblastcells (Example 13, below). The compounds have also been shown to lowerblood glucose levels in db/db mice in an insulin independent manner(FIGS. 8 and 9, Example 17; FIG. 11, Example 19).

The ability of the compounds of this invention to stimulateautophosphorylation of the insulin receptor and to stimulate the uptakeof glucose into cells which is demonstrated in the specific examples11-24 below, indicates their usefulness in the treatment and managementof subjects with diabetes. Without intending to be bound by any theory,it is believed that the compounds of the invention act directly on thekinase function of the insulin receptor and do not necessarily competewith insulin for binding at the insulin-binding site, nor do they effectactivation of the receptor by a mechanism similar to that exhibited byinsulin. Thus, they are able directly to activate the kinase toautophosphorylate, to potentiate the effect of insulin, to activate thekinase function of the receptor in phosphorylating exogenous substratesand to effect the increased uptake of glucose by adipocytes and insulinreceptor-bearing cells in general and to lower blood glucose in diabeticsubjects. Accordingly, by virtue of the activities of the compounds ofthe invention, they may be used to stimulate the kinase activity of aninsulin receptor, to enhance the activation of the insulin receptor byinsulin, to enhance the stimulation by insulin of cellular glucoseuptake, and to stimulate the uptake of glucose in diabetic subjects.Thus, the compounds of this invention are useful in the treatment ofhyperglycemia and diabetes in mammals.

One aspect of the invention is directed to a method of stimulating thekinase activity of the insulin receptor. This method comprisescontacting the insulin receptor, or the kinase portion thereof, with acompound of the invention in an amount sufficient to stimulate thekinase activity of the insulin receptor. By stimulating the kinaseactivity of the insulin receptor, both autophosphorylation as well asthe phosphorylation of exogenous substrates is enhanced. The stimulationof the kinase activity of the insulin receptor may occur either in vivoor in vitro. The method of stimulating the kinase activity of theinsulin receptor may optionally further comprise also contacting theinsulin receptor with insulin.

The compounds of the invention have been demonstrated to exhibitstimulatory activity at the insulin receptor with subsequent lowering ofcirculating glucose levels for a potential therapeutic effect indiabetes illness. Similarly, other compounds which show the same effectson the insulin receptor and, thus, on circulating glucose have thepotential to be useful for the treatment of diabetes diseases. Thecompounds of this invention can be used as a model to discover other newagents that act on the insulin receptor and thereby lower circulatinglevels of glucose in diabetic patients. The steps in a process in whichthese agents can be utilized to discover new insulin receptoragonists/activators and glucose-lowering therapeutic agents may beachieved by the following. The compounds may be utilized to validate,optimize, and standardize assays necessary for the discovery of othercompounds that:

1. Activate/stimulate the cytoplasmic kinase domain of the insulinreceptor kinase or the insulin receptor kinase;

2. Activate/stimulate the insulin receptor;

3. Stimulate glucose uptake in to cells and tissues;

4. Lower circulating glucose levels in mammals;

5. Lower circulating glucose levels in humans;

6. Inhibit lipolysis in cells and tissues;

7. Inhibit lipolysis in mammals.

These compounds can be utilized as a bench mark to discover compoundsthat show improved activity in assays that:

1. Activate/stimulate the cytoplasmic kinase domain of the insulinreceptor kinase or the insulin receptor kinase;

2. Activate/stimulate the insulin receptor;

3. Stimulate glucose uptake in to cells and tissues;

4. Lower circulating glucose levels in mammals;

5. Lower circulating glucose levels in humans;

6. Inhibit lipolysis in cells and tissues;

7. Inhibit lipolysis in mammals.

Combined with algorithms that compare structures or chemical propertiesand/or match structures or chemical properties within libraries of testcompounds, these compounds can be utilized to discover compounds thatdisplay activity in bioassays that:

1. Activate/stimulate the cytoplasmic kinase domain of the insulinreceptor kinase or the insulin receptor kinase.;

2. Activate/stimulate the insulin receptor;

3. Stimulate glucose uptake in to cells and tissues;

4. Lower circulating glucose levels in mammals;

5. Lower circulating glucose levels in humans;

6. Inhibit lipolysis in cells and tissues;

7. Inhibit lipolysis in mammals.

Combined with algorithms that compare structures and/or match structuresfor the purpose of modeling molecular interactions, these compounds canbe utilized to discover compounds that display activity in bioassaysthat:

1. Activate/stimulate the cytoplasmic kinase domain of the insulinreceptor kinase or the insulin receptor kinase;

2. Activate/stimulate the insulin receptor;

3. Stimulate glucose uptake in to cells and tissues;

4. Lower circulating glucose levels in mammals;

5. Lower circulating glucose levels in humans;

6. Inhibit lipolysis in cells and tissues;

7. Inhibit lipolysis in mammals.

Radioactive compounds of this invention can be used to diagnosediabetes, because there is a decrease in the number of insulin receptorsin patients with type II diabetes and even in patients displayingpre-diabetic risk factors such as Syndrome-X. A simple tissue biopsyfollowed by exposure of the tissue sample to these radioactive compoundscan yield a measure of the receptor count for the biopsy tissue. A lowreceptor density count can then be used to diagnose diabetes orpre-diabetes in the patient.

Radioactive compounds have a long history of use in the discovery of newdrugs. The compounds of this invention all have the potential to beeasily radiolabeled and can be used to discover other new agents thatact on the insulin receptor and thereby lower circulating levels ofglucose in diabetic patients. The process in which these agents can beutilized to discover new insulin receptor agonists/activators andglucose-lowering therapeutic agents are as follows:

Radioactive compounds of this invention can be utilized to validate,optimize, and standardize bioassays used for discovery of othercompounds that:

1. Activate/stimulate the cytoplasmic kinase domain of the insulinreceptor kinase or the insulin receptor kinase;

2. Activate/stimulate the insulin receptor;

3. Stimulate glucose uptake in to cells and tissues;

4. Lower circulating glucose levels in mammals;

5. Lower circulating glucose levels in humans;

6. Inhibit lipolysis in cells and tissues;

7. Inhibit lipolysis in mammals.

Radioactive compounds of this invention can be utilized as a bench markto discover compounds that show improved activity in bioassays that:

1. Activate/stimulate the cytoplasmic kinase domain of the insulinreceptor kinase or the insulin receptor kinase;

2. Activate/stimulate the insulin receptor;

3. Stimulate glucose uptake in to cells and tissues;

4. Lower circulating glucose levels in mammals;

5. Lower circulating glucose levels in humans;

6. Inhibit lipolysis in cells and tissues;

7. Inhibit lipolysis in mammals;

In another embodiment of the invention, the insulin receptor isactivated by contacting the insulin receptor, or the kinase portionthereof, with a compound of the invention in an mount sufficient toactivate the insulin receptor. The targeted insulin receptor mayoptionally be on the surface of a cell in a mammal. In such a case, thecontacting is effected by administering the compound, or apharmaceutical composition thereof, to the mammal. Optionally, themethod may further comprise contacting the insulin receptor withinsulin.

In an alternative embodiment, the compounds of the invention are used tostimulate the uptake of glucose into cells displaying the insulinreceptor. This method comprises contacting the cells with a compound ofthe invention, optionally in the presence of insulin, and in an amountsufficient to stimulate the uptake of glucose into the cells. Thetargeted cells may optionally be in a mammal and the step of contactingthe receptor with the compound may then be effected by administering thecompound, or pharmaceutical composition thereof, to the mammal. In oneembodiment of the method of stimulating the uptake of glucose into cellsdisplaying the insulin receptor, the cells are also contacted withexogenous insulin.

A method of treating hyperglycemia in a mammal, preferably a human, isalso contemplated by the present invention. The methods comprisesadministering a therapeutically effective amount of a compound of thisinvention, or a pharmaceutical composition thereof, to a mammal.Optionally, the method may further comprise treating the mammal with oneor more additional forms of therapy or treatment for hyperglycemia. Forinstance, one method may comprise administering exogenous insulin to themammal in addition to the compound of the invention. Alternatively, thecompounds of the invention may be administered to the mammal incombination with a non-insulin drug or other alternative treatment forhyperglycemia. The total amount of the combination of drugs administeredto the mammal must be a therapeutically effective amount, although theamounts of each of the individual drugs may by themselves be suboptimalfor therapeutic purposes.

A very dangerous side-effect of the administration of insulin isinsulin-induced hypoglycemia with the potential for coma and, possibly,death. This problem can become quite severe in diabetic patients whodevelop unpredictable responses to insulin or have hyper-variable levelsof circulating glucose. For these patients, the co-administration ofthese compounds with sub-therapeutic doses of insulin will minimize thepossibility that the diabetic patient will over-dose on insulin andsuffer from the severe consequences such as coma and death. Thesecompounds appear to be incapable of inducing hypoglycemia in thepresence or absence of insulin. They appear to increase theeffectiveness of insulin, but do not display true insulin mimeticeffects like hypoglycemia. These compounds are, thus, effective insulinsafeners.

In one embodiment of the invention, the compounds are used to treat typeI diabetes in a mammal. This method comprises administering atherapeutically effective amount of a compound of this invention, or apharmaceutical composition thereof, to the mammal. In a preferredembodiment, the mammal is a human. The method of treating type Idiabetes may optionally further comprise treating the mammal with one ormore additional therapies or treatments for type I diabetes. Forinstance, in one embodiment of the method of treating type I diabetes, acompound of the invention and insulin may both be administered to themammal. Alternatively, the additional form of treatment for type Idiabetes which is combined with administration of the compound of theinvention may be an antidiabetic agent other than insulin or anotheralternative form of treatment for type I diabetes. Again, the totalamount of the combination of antidiabetic agents administered to themammal must be a therapeutically effective amount, although the amountsof each of the individual drugs may be sub-optimal for therapeuticpurposes if those drugs were to be delivered alone to the, mammal withtype I diabetes.

In another embodiment of the invention, the compounds of the inventionare used to treat type II diabetes in a mammal. This method comprisesadministering a therapeutically effective amount of a compound of thisinvention, or a pharmaceutical composition thereof, to the mammal.Again, the preferred subject is a human.

Again, like the other treatment methods of the invention, this methodmay further comprise treating the mammal with one or more additionalforms of therapy for type II diabetes, such as administering insulin tothe mammal. The insulin is delivered to the mammal in an amount which istherapeutically effective when used in conjunction with a compound ofthe invention. This therapeutically effective amount of insulin whenused in combination with a compound of the invention may be less thanthe amount of insulin which would be therapeutically effective ifdelivered to the mammal alone. It is understood that the insulin whichis administered in any of the treatments of the present invention mayeither be isolated from a natural source or be recombinant. In addition,an insulin analog may be substituted for insulin in any of thetreatments of the present invention.

Use of the compounds of the invention for treating type II diabetes bycombination therapy may also comprise the administration of the compoundof the invention to the mammal in combination with a non-insulin,antidiabetic agent or other treatment for type II diabetes. Forinstance, the antidiabetic drug which is administered to the mammal incombination with a compound of the invention may optionally be athiazolidinedione, such as troglitazone, or a sulfonylurea. The totalamount of the combination of drugs (invention compound plus insulin,and/or other antidiabetic drug) administered to the mammal for thetreatment of type II diabetes must be a therapeutically effectiveamount, although the amount of each of the individual drugs used in thecombination therapy may be suboptimal for therapeutic purposes if thatdrug were to be delivered alone to the mammal with type II diabetes.

The compounds of this invention are thus used to enhance glucose uptakein patients which require such treatment. The method of treatmentcomprises the administration parenterally, and orally, of an effectivequantity of the chosen compound of the invention, preferably dispersedin a pharmaceutical carrier. Dosage units of the active ingredient aregenerally selected from the range of 0.01 to 1000 mg/kg, preferably 0.01to 100 mg/kg and more preferably 0.1 to 50 mg/kg, but will be readilydetermined by one skilled in the art depending upon the route ofadministration, age and condition of the patient. The compounds of theinvention are most preferably administered in a dosage unit of 1 to 10mg/kg. These dosage units may be administered one to ten times daily foracute or chronic disease. No unacceptable toxicological effects areexpected when compounds of the invention are administered in accordancewith the present invention.

The invention compounds may be administered by any route suitable to thesubject being treated and the nature of the subject's condition. Routesof administration include, but are not limited to, administration byinjection, including intravenous, intraperitoneal, intramuscular, andsubcutaneous injection, by transmucosal or transdermal delivery, throughtopical applications, nasal spray, suppository and the like or may beadministered orally. Formulations may optionally be liposomalformulations, emulsions, formulations designed to administer the drugacross mucosal membranes or transdermal formulations. Suitableformulations for each of these methods of administration may be found,for example, in Remington's Pharmaceutical Sciences, latest edition,Mack Publishing Company, Easton, Pa.

Methods, uses, activities, administration, and pharmaceuticalcompositions delineated above for the compounds of the invention arepreferred for those compounds in which K═O.

(d) EXAMPLES

The Examples which follow serve to illustrate this invention. TheExamples are in no way intended to limit the scope of this invention,but are provided to show how to make and use the compounds of thisinvention.

The compounds of formula I may be prepared by:

(i) intermolecular or intramolecular condensation of a compound of theformula

 with a compound of the formula

 or the addition of one of these compounds to the other, where the aminogroups of the compounds are optionally in a protected form, with anactivated bifunctional reagent that provides the group K═C—, where K hasthe above meaning; where R¹, R², R³ , R⁴, R⁵, R⁶, x, and Y are definedaccording to the first aspect of the Summary of the Invention, X may beR³ or R⁴, and WZ may be S═C═ or O═C═, or W and Z may be R³ or R⁴;

to form a linker between the two compounds and prepare a compound ofFormula I;

(ii) chemical elaboration of one or more substituents R³, R², R⁵, and R⁶or Y, where said substituent is convertible into another substituent;

(iii) introduction of a substituent R¹, R², R⁵, and R⁶ or Y into one orboth of the naphthalene rings;

(iv) deprotection;

(v) elaboration of the linker to convert said linker into anotherlinker;

(vi) salt formation or interconversion;

(vii) ester hydrolysis;

(viii) liberation of a free acid or base of a compound of claim 1; or

(ix) stereoisomer separation or synthesis.

Details are apparent from the following table, which shows typicalreactions for steps (i) through (viii).

In a condensation reaction, a simple substance, such as. water, isreleased by the combination of two or more molecules. The condensationreaction may occur upon addition of any of a number of startingmaterials utilized in organic syntheses, such as dibromoethane anddiiodopropane, at a temperature between 50 and 125 C. Should R³ and R⁴in the formulae above together be —(CH₂)₂, —(CH₂)₃—, or (CH₂)₄—, thenthe condensation is intramolecular (see Reaction Scheme VI).

Chemical elaboration of one or more substituents R₁, R², R⁵, and R⁶ or Yvia the conversion of one such substituent into another substituent maybe accomplished via hydrolysis, salt formation, acidification,alkylation, esterification, oxidation, or reduction. In hydrolysis, anester or amide compound is dissociated by reaction with water.Hydrolysis is catalyzed by acid or base, and hydrolysis of an amidegenerally requires more vigorous conditions (for example, a higherconcentration of sulfuric acid at 1 to 100 C for 1 to hours) than thoserequired for the hydrolysis of esters. Hydrolysis reactions can also becarried out with aqueous hydrochloric acid at 100 to 150 C. and mayrequire as long as 18 hours.

In salt formation, a free acid is converted into a salt via addition ofa basic reagent, such as aqueous sodium hydroxide or triethanolamine,that replaces all or part of the hydrogen ions of the acid with one ormore cations of a base. The conversion of a compound into itscorresponding acid addition salt is accomplished via treatment with astoichiometric amount of an appropriate acid, such as hydrochloric acid.Typically, the free base is dissolved in a polar organic solvent, suchas methanol or ethanol, and the acid is added in methanol or ethanol.The temperature is maintained at 0 to 50 C. The corresponding saltprecipitates spontaneously or can be brought out of solution with a lesspolar solvent. In acidification, a chemical compound is converted intoan acid.

In alkylation, an alkyl group is added to or substituted in a compound.Alkylation is carried out in a suitable solvent, such as acetonitrile,DMF, or THF, at 0 to 160 C., typically at approximately 25 C. to reflux,and requires some 1 to 18 hours. Finally, an esterification reactionresults in the formation of at least one ester product. In brief, thecompound is reacted with from 1.0 to 5.0, preferable 2.0, molarequivalents of an alkanol, a thiol or ammonia, a monoalkylamine, ordialkylamine, or a heterocyclic aminoalkanol, optionally in the presenceof from 1.0 to 1.5, preferably 1.25, molar equivalents of a tertiaryorganic base such as 4-dimethylaminopyridine or, preferably,triethylamine, in an organic solvent such as dioxane, tetrahydrofuran,or, preferably, dichloromethane. The reaction takes place at −10 to 50C., preferably at ambient temperature, for 1 to 24 hours, preferably 4hours.

Certain compounds of formula I can be prepared via acid addition.Furthermore, the compounds of formula I may be prepared by modifying K(where K has the above meaning), for example, by alkylating K, followedby amino substitution, in which an amino group replaces, for example, aleaving group such as the S-methyl group.

In those cases in which protective groups may be introduced and finallyremoved, suitable protective groups for amino, hydroxy, carboxyl groupsare as described in Greene, et al., Protective Groups in OrganicSynthesis, Second Edition, John Wiley and Sons, New York, 1991.Activation of carboxylic acids can be achieved by using a number ofdifferent reagents as described in Larock, Comprehensive OrganicTransformations, VCH Publishers, New York, 1989.

Example 1

Compounds 3, 4, 8-16, and 19-93 were prepared according to ReactionScheme I and Reaction Scheme II.

7-{[(7-sulfo-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic aciddisodium salt (Compound 3)

To 5.25 g (0.021 mol) of 7-amino-naphthalene-2-sulfonic acid suspendedin 80 mL of water was added a solution of 6.5 mL of 10 N aqueous NaOH(0.065 mol) diluted to 30 mL with water and a solution of 3.20 g (0.011mol) of triphosgene in 30 mL of THF portionwise, and alternating suchthat the pH of the reaction was maintained above 8. After the reactionwas complete by TLC (6:2:1 ethyl acetate:isopropanol:water) the pH waslowered to 1 with aqueous HCl and the volatiles were removed by rotaryevaporation. The solid product was collected by vacuum filtration andwas washed with water. This afforded 3.41 g of compound 3.

4-hydroxy-7-{[(5-hydroxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid disodium salt (Compound 4)

To 10.77 g (0.045 moles) of 7-amino-4-hydroxynaphthalene-2-sulfonic aciddissolved in 45 mL of 1 N aqueous NaOH and 50 mL of water was added 3.70g (0.045 moles) of sodium acetate. The pH of the solution was above 9.The reaction was cooled to under 5° C. in an ice-water bath. Then, 2.23g (0.045 mole) of triphosgene dissolved in mL of THF was added in threeportions. The pH of the reaction fell to 4-5 and was readjusted to 7-8by the dropwise addition of 1N aqueous NaOH. TLC (6:2:1 ethylacetate:isopropanol:water) indicated the reaction was incomplete.Another 2.20 grams (0.045 moles) of triphosgene in 10 mL of THF wasadded portionwise with the pH kept above 7 by the addition of 1N aqueousNaOH. When the reaction was judged complete by TLC, the pH was loweredto 1 with aqueous HCl and the volatiles were removed by rotaryevaporation. The solid product was collected by vacuum filtration. Thisafforded 10.85 g of compound 4.

4-methylphenyl 3-aminobenzenesulfonate (Compound 5)

To 50 g (0.463 mole) of p-cresol (4-methylphenol) and 37 mL (0.458 mole)of pyridine dissolved in 250 mL of chloroform was added 50 g (0.226mole) of 3-nitrobenzenesulfonyl chloride. The reaction was allowed tostir at ambient temperature. After 2 hours, the reaction was judgedcomplete by TLC and the volatiles were removed by rotary evaporation.The resulting residue was treated with 400 mL of 0.5 M sodiumbicarbonate. The insoluble product was collected and washed with sodiumbicarbonate (2 times, 200 mL each) and water (2 times, 300 mL each). Thesolid was then treated with methanol (200 mL) followed by water (200mL). The solid was collected by vacuum filtration and washed with water.This solid was treated with 170 g (0.897 mole) of tin (II) chloridedissolved in 250 mL of concentrated HCl. The reaction was allowed tostir at ambient temperature for 40 hours. TLC indicated that thereaction was incomplete, so the reaction was heated at 50 C. for 27hours. The solid precipitate was collected by vacuum filtration and waswashed with 6N HCl. The solid was extracted with ethyl acetate and,water. The ethyl acetate layer was washed with brine, dried withmagnesium sulfate, filtered, and the volatiles removed by rotaryevaporation to yield 42.5 g of compound as an oil that solidified onstanding.

N-[7-(chlorosulfonyl)(2-naphthyl)]{[7-(chlorosulfonyl)(2-naphthyl)]amino}carboxamide(Compound 6)

To 2.35 g (4.86 mmol) of compound 3 was added 116 mL of sulfolane, 25 mLof acetonitrile, 31 mL of phosphorus oxychloride, and 1 mL ofdimethylacetamide. The reaction was allowed to stir for 72 hours atambient temperature. This produced a nearly clear solution. The reactionwas poured onto 1.5 L of ice and the flask placed in an ice bath. Afterall the ice had melted, the solid was collected by vacuum filtration andwas washed with water. The solid was dried under high vacuum for 24hours. This provided 2.56 g of compound 6.

7-{[(7-(chlorosulfonyl)-5-hydroxy(2-naphthyl))amino]carbonylamino}-4-hydroxynaphthalene-2-sulfonylchloride (Compound 7)

To 500 mg (0.912 mmol) of compound 4 suspended in 8 mL of phosphorousoxychloride was added 25 mL of 1:1 (v:v) sulfolane:acetonitrile and 0.5mL of dimethylacetamide. The reaction mixture was allowed to stir atambient temperature for 16 hours. The reaction became a clear solutionwhich was poured onto 500 mL of ice. The ice mixture was placed in anice bath and allowed to warm to room temperature. The resulting solidwas collected by vacuum filtration and was washed with water. The solidwas dried under high vacuum for 24 hours. This provided 412 mg ofcompound 7.

7-[({7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}amino)carbonylamino]naphthalene-2-sulfonicacid (compound 8) and 4-methylphenyl3-[({7-[(N-{7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate(Compound 9)

To 250 mg (0.95 mmol) 4-methylphenyl 3-aminobenzenesulfonate (compound5). Dissolved in 40 mL of pyridine was added a solution of 250 mg (0.49mmol) of compound 6 in 5 mL of THF. The reaction was allowed to stir atambient temperature for 3 hours. Then, the reaction was extracted withethyl acetate and 1 N HCl. The ethyl acetate layer was dried withmagnesium sulfate, filtered, and the volatiles were removed by rotaryevaporation. The products were purified by reverse-phase (RP) HPLC (C18,30×250 mm column) using trifluoroacetic acid (TFA) buffer system (BufferA: 5% acetonitrile, 95% water, 0.05% TFA; Buffer B: 95% acetonitrile, 5%water, 0.05% TFA; 35 mL/min, 0-100% B in 45 min.). Fractions containingthe earlier eluting compound were combined and lyophilized to provide 9mg of compound 8. Fractions containing the later eluting compound werecombined and lyophilized to provide 51 mg of compound 9.

4-hydroxy-7-[({5-hydroxy-7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl](2-naphthyl)}amino)carbonylamino]naphthalene-2-sulfonicacid (compound 10) and 4-methylphenyl3-[({4-hydroxy-7-[(N-{5-hydroxy-7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl](2-naphthyl)}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate(Compound 11)

To 250 mg (0.95 mmol) 4-methylphenyl 3-aminobenzenesulfonate (compound5) dissolved in 40 mL of pyridine was added a solution of 265 mg (0.49mole) of compound 7 in 5 mL of THF. The reaction was allowed to stir atambient temperature for 3 hours. Then, the reaction was extracted withethyl acetate and 1 N HCl. The ethyl acetate layer was dried withmagnesium sulfate, filtered, and the volatiles were removed by rotaryevaporation. The products were purified by reverse-phase HPLC (C18,30×250 mm column) using trifluoroacetic acid (TFA) buffer system (BufferA: 5% acetonitrile, 95% water, 0.05% TFA; Buffer B: 95% acetonitrile, 5%water, 0.05% TFA; 35 mL/min, 0-100% B in 60 min.). Fractions containingthe earlier eluting compound were combined and lyophilized to provide 15mg of compound 10. Fractions containing the later eluting compound werecombined and lyophilized to provide 65 mg of compound 11.

7-{[(7-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonicacid disodium salt (Compound 12)

To 8 mg ( 0.011 mmol) of compound 8 was added 2 mL of 5 N NaOH. Thereaction was allowed to stir at ambient temperature for 6 hours. Thereaction was acidified with 6 N HCl and the solution was added to asmall reverse-phase (C18) solid phase extraction column. The column waswashed with H₂O followed by 50:50 (v:v) CH₃CN:H₂O to elute the product.This provided 5 mg of compound 12.

3-{[(7-{[N-(7-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid disodium salt (Compound 13)

To 35 mg ( 0.036 mmol) of compound 9 was added 2 mL of 5 N NaOH. Thereaction was allowed to stir at ambient temperature for 6 hours. Thereaction was acidified with 6 N HCl and the solution was added to asmall reverse-phase (C18) solid phase extraction column. The column waswashed with H₂O followed by 50:50 (v:v) CH₃CN:H₂O to elute the product.This provided 26 mg of compound 13.

4-hydroxy-7-{[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid (Compound 14)

This compound was prepared from compound 10 according to the proceduredescribed for the synthesis of compound 12.

3-{[(4-hydroxy-7-{[N-(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulonicacid (Compound 15)

This compound was prepared from compound 11 according to the proceduredescribed for the synthesis of compound 12.

3-bromo-7-{[(6-bromo-5-hydroxy-7-sulfo(2-naphthyl))amino]carbonylamino}-4-hydroxynaphthalene-2-sulfonicacid disodium salt (Compound 16)

To a solution of 123 mg (0.22 mmol) of compound 4 in 1 mL of water and 2mL of dioxane was added 300 μL of a 1 M solution of bromine in carbontetrachloride. After 1 hour, another 200 μL of the bromine solution wasadded to the reaction. After another hour, an additional 150 μL of thebromine solution was added to the reaction. After 30 minutes, thereaction was poured into 100 mL of THF to produce a brown precipitatethat was collected by vacuum filtration. The desired product waspurified by silica gel column chromatography (5:2:1 ethylacetate:isopropanol:water) to provide 17 mg of compound 16.

Example 2

An alternative synthetic method for the compounds of the invention isdescribed in Reaction Scheme II. This method was useful for thesynthesis of larger quantities of the desired products and isillustrated by the synthesis of compound 15.

6-(acetylamino)-3-(chlorosulfonyl)naphthyl acetate (Compound 17)

To 50 g (0.209 mmol) of compound 2 was added 100 mL of acetic anhydrideand 100 mL of pyridine. The reaction was allowed to stir at ambienttemperature for 16 hours. The solid precipitate was collected by vacuumfiltration. This provided 83 g of product that was dried under highvacuum. To this solid was added 260 mL of phosphorous oxychloride. Thissuspension was allowed to stir at ambient temperature for 16 hours.Then, the dark solution was poured onto 3 L of ice. After all the icehad melted (about 3 hours), the solid was collected by vacuum filtrationand washed with water. The solid was dried in vacuo to provide 50 g ofcompound 17.

3-{[(7-amino-4-hydroxy-2-naphthyl)sulfonyl]amino}benzenesulfonic acidsodium salt (Compound 18)

To 25 g (0.095 mol) of compound 5 dissolved in 200 mL of THF was added8.1 mL of pyridine. Then, 36.3 g (0.106 mol) of compound 17 was addedfollowed by an additional 200 mL of THF. The reaction was allowed tostir at ambient temperature for 16 hours. Then, the volatiles wereremoved and the resulting residue was partitioned between ethyl acetateand 1N HCl. The ethyl acetate layer was washed with brine, treated withmagnesium sulfate, filtered and the volatiles removed by rotaryevaporation. The resulting solid was dissolved in 220 mL of 5N NaOH and30 mL of dioxane. The solution was heated at 80 C. for hours. Then, thepH of the solution was lowered to 1 with concentrated HCl. The solid wascollected by vacuum filtration and then dissolved in 200 mL of waterwith 20 mL of 5 N NaOH. The solution was heated to give a hazy solutionthat was filtered hot to produce a clear solution. The solution wasdiluted with water to 1.5 L and then the pH was adjusted to 1 with 6 NHCl. A solid precipitate formed. The suspension was cooled in therefrigerator overnight and the solid was collected by vacuum filtrationto furnish, after drying, 21 g of compound 18.

3-{[(4-hydroxy-7-{[N-5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]-amino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid disodium salt (Compound 15)

To 17.7 g (0.045 mol) of compound 18 was added 250 mL of acetate buffer(1M NaOAc/HOAc, pH 4.6) and 50 mL of THF. This formed a dark brownsolution. A solution of 4.4 g (mmol) of triphosgene in 40 mL of THF wasadded dropwise to the above solution over a period of 3 hours and 10min. HPLC indicated the reaction was complete. The reaction wasacidified with concentrated HCl (15 mL). All the volatiles were removedby rotary evaporation, and the resulting goo was stripped fromacetonitrile, acetonitrile/heptane, and then heptane. The resultingsolid was dissolved in 140 mL of water with heating. Then, 250 mL ofbrine was added. A solid precipitate formed. The flask was kept at 4 C.for 15 hours, and then the solid was collected by vacuum filtration. Thesolid was washed with a small amount of ice-cold 3:1 water:brine. Afterdrying under high vacuum, this provided 19.2 g of compound 15.

The following compounds (Table 1) were prepared according to the samegeneral procedures as those outlined in Reaction Scheme I or ReactionScheme II and detailed in the above procedures for the synthesis ofcompounds 3-18. The variety of molecules that were used in place ofcompound 5 for the synthesis of compounds 19-91 were either purchasedfrom commercial suppliers or were prepared by standard methods known tothose skilled in the art. Compounds in Table 1 with acidic groups areshown in the free acid form.

TABLE 1

Compound R⁰ R⁵ R⁶ R⁰⁰ Y 3 OH H H OH H 4 OH OH OH OH H 8 OH H H

H 9

H H

H 10 OH OH OH

H 11

OH OH

H 12 OH H H

H 13

H H

H 14 OH OH OH

H 15

OH OH

H 16 OH OH OH OH Br 19 OH H H

H 20 OH H H

H 21

H H

H 22

H H

H 23

H H

H 24

H H

H 25

H H

H 26 OH OH OH

H 27

OH OH

H 28 OH OH OH

H 29

OH OH

H 30 OH H H

H 31

H H

H 32 OH H H

H 33

H H

H 34 OH H H

H 35

H H

H 36 OH H H

H 37 OH H H

H 38

H H

H 39

H H

H 40

H H

H 41

OH OH

H 42

OH OH

H 43

OH OH

H 44

H H

H 45

H H

H 46

OH OH

H 47

OH OH

H 48

H H

H 49

OH OH

H 50

OH OH

H 51

OH OH

H 52

OH OH

H 53

H H

H 54

H H

H 55

H H

H 56

H H

H 57

H H

H 58

H H

H 59

OH OH

H 60

OH OH

H 61

H H

H 62

H H

H 63

H H

H 64

H H

H 65

H H

H 66

H H

H 67

H H

H 68

OH OH

H 69

OH OH

H 70

H H

H 71

H H

H 72 OH H H

H 73 OH H H

H 74

H H

H 75

H H

H 76

H H

H 77

H H

H 78 OH H H

H 79

H H

H 80

H H

H 81

H H

H 82

OH OH

H 83

H H

H 84

H H

H 85

H H

H 86

H H

H 87

H H

H 88

OH OH

H 89 NH₂ H H NH₂ H 90 OH H H

H 91

H H

H 92 OH OH OH

H 93

OH OH

H

Example 3

The compounds 94-102 were synthesized according to the proceduresoutlined in Reaction Scheme III.

7-[({5-[(ethoxycarbonyl)methoxy]-7-sulfo(2-naphthyl)}amino)carbonylamino]-4-hydroxynaphthalene-2-sulfonicacid, diammonium salt (Compound 94) and4-[(ethoxycarbonyl)methoxy]-7-[((5-[(ethoxycarbonyl)methoxy]-7-sulfo(2-naphthyl)}amino)-carbonylamino]naphthalene-2-sulfonicacid, diammonium salt (Compound 95)

To 275 mg (0.5 mmol) of compound 4 suspended in 50 mL of ethanol wasadded 2.5 mL of a 0.2M solution of sodium ethoxide in ethanol. Then, 56μL (0.5 mmol) of ethyl bromoacetate was added. The mixture was stirredand refluxed for 2 hours. Then, the solid was removed by filtration andthe volatiles were removed from the filtrate to provide 220 mg of crudeproduct. The products were purified by reverse-phase HPLC (C18, 20×250mm column) using ammonium acetate (NH₄OAc) buffer system (Buffer A: 5%acetonitrile, 95% water, 50 mM NH₄OAc; Buffer B: 70% acetonitrile, 30%water, 15 mM NH₄OAc; 15 mL/min, 0-100% B in 30 min.). Fractionscontaining the earlier eluting compound were combined and lyophilized toprovide 60 mg of compound 94. Fractions containing the later elutingcompound were combined and lyophilized to provide 32 mg of compound 95.

2-(6-{[N-(5-hydroxy-7-sulfo(2-naphthyl)carbamoyl]amino}-3-sulfonaphthyloxy)aceticacid disodium salt (Compound 96)

To 25 mg (0.04 mmol) of compound 94 was added 1 ML of 5 N NaOH. Thesolution was allowed to stir for 18 hours at ambient temperature. The pHwas lowered to 1 with aqueous HCl and the resulting solid was collectedby vacuum filtration to afford 16 mg of compound 96.

2-[6-({N-[5-(carboxymethoxy)-7-sulo(2-naphthyl)]carbamoyl}amino)-3-sulfonaphthyloxy]aceticacid disodium salt (Compound 97)

Compound 95 was treated as above for compound 94 to provide 11 mg ofcompound 97.

The following compounds (Table 2) were prepared according to the samegeneral procedures as those outlined in Reaction Scheme III and detailedin the above procedures for the synthesis of compounds 94-97. Thevariety of molecules that were used in place of ethyl bromoacetate forthe synthesis of compounds 98-102 were either purchased from commercialsuppliers or were prepared by standard methods known to those skilled inthe art. All of the compounds in Table 2 are shown in the free acidform.

TABLE 2

Com- pound R⁰ R⁵ R⁶ R⁰⁰ 94 OH OH

OH 95 OH

OH 96 OH OH

OH 97 OH

OH 98 OH OH OCH₃ OH 99 OH OCH₃ OCH₃ OH 100 OH

OH 101 OH OH

OH 102 OH

OH

Example 4

The compounds 103-105 were synthesized according to the procedureoutlined in Reaction Scheme IV.

3-{methyl[(7-{[(7-{[methyl(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]-carbonylamino}(2-naphthyl))sulfonyl]amino}benzenesulfonicacid dipotassium salt (Compound 103)

To 59 mg (0.071 mmol) of compound 13 was added 2 mL of DMF. Then, 38 mgof potassium carbonate was added. The stirred suspension was heated at70 C. in an oil bath. Then, 36 μL of iodomethane was added. The reactionwas heated for 3 hours, and then the volatiles were removed by rotaryevaporation. The resulting solid was dissolved in water and added to aC18 solid phase extraction column. The column was washed with 9 columnvolumes of water. The product was eluted with 80% acetonitrile tofurnish, after evaporation of the volatiles, 55 mg of compound 103.

Compounds 104 and 105 (Table 3) were prepared by the same generalprocedure as indicated above for the synthesis of compound 103 exceptthat allyl bromide was used in place of methyl iodide and compound 48was used in place of compound 13. Compounds in Table 3 with acidicgroups are shown in the free acid form.

TABLE 3

Compound # R^(I) R^(II) R^(III) R^(IV) 103

CH₃ CH₃

104

CH₂CH═CH₂ CH₂CH═CH₂

105

CH₂CH═CH₂ CH₂CH═CH₂

Example 5

The compounds 112-114 and 116 were synthesized according to proceduresoutlined in Reaction Scheme V.

7-(fluoren-9-yloxycarbonylamino)naphthalene-2-carboxylic acid (107)

To 0.504 g (2.70 mmol) of 7-aminonaphthalene-2-carboxylic acid (106;prepared according to the procedure in Harrison, H. A. and Royle, F. A.J. Chem. Soc., 1926, 84) was added 10 mL of dioxane, 5 mL of 10% sodiumcarbonate, and 35 mL of water. To this clear solution was added 0.786 g(2.97 mmol) of 9-fluorenylmethyl chloroformate, portionwise, over 15minutes. After 3 hours, the reaction was acidified with 1N HCl and theresulting white precipitate was collected by vacuum filtration. Thissolid was suspended in diethyl ether and stirred to give a fineprecipitate that was collected by vacuum filtration. This provided 0.948g of compound 107.

4-methylphenyl3-{[7-(fluoren-9-yloxycarbonylamino)-2-naphthyl]carbonylamino}benzenesulfonate(Compound 108)

To 104 mg (0.026 mmol) of compound 107 was added 6.5 mL of chloroform,1.3 mL of thionyl chloride, and 100 μL of pyridine. The reaction wasallowed to stir at ambient temperature for 3 hours and then allvolatiles were removed in vacuo. The residue was stripped in vacuo fromchloroform twice. To the resulting solid, suspended in 15 mL ofchloroform was added 74 mg (0.028 mmol) of 4-methylphenyl3-aminobenzenesulfonate (compound 5) and 27 μL (0.033 mmol) of pyridineas a solution in 1.5 mL of chloroform. The reaction was allowed to stirat ambient temperature for 16 hours, after which it was partitionedbetween ethyl acetate and 1 N HCl (aqueous). The organic layer was dried(magnesium sulfate), filtered, and the volatiles were removed in vacuo.The resulting residue was treated with diethyl ether. and the solidprecipitate collected by vacuum filtration. This provided 110 mg ofcompound 108.

4-methylphenyl 3-{[(7-amino-2-naphthyl)sulfonyl]amino}benzenesulfonate(Compound 110)

To 104 mg (0.16 mmol) of compound 108 was added 9 mL of THF and 360 μLof piperidine. The resulting clear solution was allowed to stir for 6hours. Then, the reaction was extracted with ethyl acetate and 1N HCl(aqueous). The dried organic layer (magnesium sulfate) was filtered andthe volatiles removed in vacuo. The resulting residue was dissolved indichloromethane and 3 mL of 1 N HCl in diethyl ether and 50 mL ofdiethyl ether were added to form a precipitate that was collected bycentrifugation. After drying, this provided 75 mg of compound 110 as thehydrochloride salt.

4-methylphenyl3-[({7-[(N-{7-[{(3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate(Compound 112)

To 75 mg (0.17 mmol) of compound 110 dissolved in 3 mL of THF and 1 mLof water was added, portionwise and alternating, a solution of 280 μL of5 N NaOH (aqueous) in 1 mL of water followed by a solution of 54 mg(0.18 mmol) of triphosgene in 1 mL of THF. The volatiles were removeduntil a solid precipitate formed and a clear solution. The solution wasdecanted and the solid was dissolved in dichloromethane, and evaporated3 times. This produced a dichloromethane insoluble product that wascollected by vacuum filtration to provide 24 mg of compound 112.

3-{[(7-{[N-(7-{[(3-sulophenyl)amino]sulfonyl)-2-naphthyl)carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid disodium salt (Compound 114)

To 20 mg (0.02 mole) of compound 112 was added 1.5 mL of 1.37 M sodiummethoxide in methanol, 1 mL of water, and 0.5 mL of THF. The resultingsolution was allowed to stir at ambient temperature for 2 days. Thereaction was acidified with 1 N HCl (aqueous) and the organic volatilesremoved in vacuo. The solid precipitate was collected by vacuumfiltration to afford 15 mg of compound 114.

methyl3-({7-[fluoren-9-ylmethoxy)carbonylamino]-2-naphthyl}carbonylamino)benzoate(Compound 109)

To 305 mg (0.75 mmol) of compound 107 was added. 10 mL of chloroform,3.5 mL of thionyl chloride, and 180 μL of pyridine. The reaction wasallowed to stir at ambient temperature for three hours, followed byremoval of volatiles by rotary evaporation. The resulting residue wasstripped two times from chloroform. Then, 50 mL of chloroform, 124 mg(0.82 mmol) of methyl-3-aminobenzoate, and 100 μL of pyridine wereadded. The reaction was allowed to stir at ambient temperature for 16hours. The reaction was extracted twice with 1N HCl (aqueous) and oncewith water. The dried organic layer (magnesium sulfate) was filtered andthe volatiles removed by rotary evaporation. The resulting residue wastreated with methanol to form a solid precipitate that was collected byvacuum filtration. This afforded 380 mg of compound 109.

methyl 3-[(7-amino-2-naphthyl)carbonylamino]benzoate (Compound 111)

To 380 mg (0.70 mmol) of compound 109 was added 30 mL of dichloromethane, 3 mL of THF and 1 mL of piperidine. The resulting clear solution wasallowed to stir for 3 hours. Then, the reaction was extracted with ethylacetate and 1N HCl (aqueous). The dried organic layer (magnesiumsulfate) was filtered and the volatiles removed in vacuo. The resultingresidue was dissolved in dichloromethane and 3 mL of 1 N HCl in diethylether and 50 mL of diethyl ether were added to form a precipitate thatwas collected by vacuum filtration. After drying, this provided 212 mgof compound 111 as the hydrochloride salt.

methyl3-[(7-{[N-(7-{N-[3-(methoxycarbonyl)phenyl]carbamoyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)carbonylamino]benzoate(Compound 113)

To 21 mg (0.07 mmol) of compound 111 dissolved in 2 mL of THF and 1 mLof water was added, portionwise and alternating, a solution of 112 μL of5 N NaOH (aqueous) in 1 mL of water followed by a solution of 28 mg(0.10 mmol) of triphosgene in 1 mL of THF. The reaction was acidifiedwith 1N HCl and the volatiles were removed until a solid precipitateformed and a clear solution. The solid was collected by vacuumfiltration to provide 21 mg of compound 113.

3-[(7-amino-2-naphthyl)carbonylamino]benzoic acid (Compound 115)

To 51 mg (0.16 mmol) of compound 111 was added 1 mL of methanol, 1 mL ofwater, and 800 μL of 1N lithium hydroxide (aqueous). The reaction wasallowed to stir for 16 hours at ambient temperature. The pH of thesolution was lowered to 3 with 1N HCl (aqueous) and the volatilesremoved in vacuo. The resulting solid was suspended in water andcollected by vacuum filtration. This provided 16 mg of compound 115.

3-({7-[(N-(7-[N-(3-carboxyphenyl)carbamoyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}carbonylamino)benzoicacid (Compound 116)

To 10 mg (0.03 mmol) of compound 115 dissolved in 2 mL of THF and 1 mLof water was added, portionwise and alternating, a solution of 40 μL of5 N NaOH (aqueous) in 0.2 mL of water followed by a solution of 4 mg(0.02 mmol) of triphosgene in 0.2 mL of THF. The reaction was acidifiedwith 1N HCl and the volatiles were removed in vacuo. The solid wascollected by vacuum filtration to provide 9 mg of compound 116.

The compounds 112-114 and 116 that were prepared by the proceduresdescribed above are indicated in Table 4. Compounds with acidic groupsare shown in the free acid form.

TABLE 4

Compound R 112 —SO₃PhCH₃ 113 —CO₂CH₃ 114 —SO₃H 116 —CO₂H

Example 6

The compounds 122 and 123 were synthesized according to the proceduresoutlined in Reaction Scheme VI.

7-({2-[(7-sulfo-2-naphthyl)amino]ethyl}amino)naphthalene-2-sulfonicacid, dipotassium salt (Compound 117)

To 2.04 g ( 9.15 mmol) of 7-amino-2-naphthalene sulfonic acid(compound 1) was added 50 mL of dry DMF. The stirred suspension washeated at 110 C. and then 1.4 g of potassium carbonate was addedfollowed by 400 μL (4.6 mmol) of 1,2-dibromoethane. The reaction waskept at 110 C for 18 hours, and at 80 C. for an additional 18 hours.Then, the reaction was allowed to cool to ambient temperature. Theresulting insoluble precipitate was collected by vacuum filtration andwashed with methanol. The crude product was purified by silica gelcolumn chromatography (6:2:1 ethyl acetate:isopropanol:water). Thisprovided 596 mg of compound 117.

7-((fluoren-9-ylmethoxy)-N-{2-[(fluoren-9-ylmethoxy)-N-(7-sulfo(2-naphthyl))carbonylamino]ethyl}carbonylamino)naphthalene-2-sulfonicacid, disodium salt (Compound 118)

To 259 mg (0.47 mmol)of compound 117 was added 25 mL of water and 207 mgof sodium carbonate. Then, 20 mL of dioxane was added. To this stirred,hazy solution was added 290 mg (1.1 mmol) of 9-fluorenylmethylchloroformate (FMOC-Cl) portionwise. After 2 hours, another 260 mg (1.0mmol) of FMOC-Cl was added and the reaction was allowed to stir atambient temperature for 12 hours. The volatiles were removed by rotaryevaporation and the resulting solid was dissolved in water andlyophilized. This crude product was used without further purification.

methyl5-[({7-[(fluoren-9-ylmethoxy)-N-(2-{(fluoren-9-ylmethoxy)-N-[7-({[4-hydroxy-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]carbonylamino}ethyl)carbonylamino](2-naphthyl))sulfonyl)amino]-2-hydroxybenzoate(Compound 119)

To all of the product compound 118 was added 15 mL of phosphorousoxychloride. This suspension was allowed to stir at ambient temperaturefor 24 hours. The yellow suspension was poured onto 700 mL of ice. Afterthe ice melted, the yellow solid was collected by vacuum filtration andwas washed with water.

The solid was dried in vacuo, overnight, to provide.194 mg of theintermediate disulfonyl chloride. To this solid was added 3 mL of THFfollowed by a solution of 72 mg (0.43 mmol) of methyl5-amino-2-hydroxybenzoate and 41 μL of pyridine in 1.5 mL of THF. Thereaction was allowed to stir at ambient temperature for 12 hours. Thereaction was partitioned between ethyl acetate and 1N HCl. The organiclayer was dried (MgSO₄), filtered, and the volatiles removed by rotaryevaporation. The resulting solid was purified by silica gel columnchromatography eluting with 0.5% methanol in dichloromethane. Thisprovided 116 mg of compound 119.

methyl2-hydroxy-5-[({7-[(2-{[7-({[4-hydroxy-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}ethyl)amino](2-naphthyl)}sulfonyl)amino]benzoate(Compound 120)

To 99 mg (0.08 mmol) of compound 119 was added 9 mL of a 5% (v/v)solution of piperidine in THF. The suspension was allowed to stir atambient temperature for 24 hours. The reaction was partitioned betweenethyl acetate and 1N HCl. The organic layer was dried (MgSO₄), filtered,and the volatiles removed by rotary evaporation. The product waspurified by silica gel column chromatography eluting with 1% methanol indichloromethane and then 2% methanol in dichloromethane. This provided64 mg of compound 120.

5-({[7-({2-[(7-{[(3-carboxy-4-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))amino]-ethyl}amino)(2-naphthyl)]sulfonyl}amino)-2-hydroxybenzoicacid (Compound 121)

To 64 mg (0.08 mmol) of compound 120 was added 20 mL of saturated sodiumcarbonate, 76 mg of sodium hydrosulfite (Na₂S₂O₄), and 2 mL of DMF. Thereaction was allowed to stir at ambient temperature for 22 hours. Then,the reaction was extracted with ethyl acetate and 1 N HCl. The organiclayer was dried (MgSO₄), filtered, and the volatiles removed by rotaryevaporation. The product was purified by reverse-phase BPLC (C18, 250×20mm column) using trifluoroacetic acid (TFA) buffer system (Buffer A: 5%acetonitrile, 95% water, 0.05% TFA; Buffer B: 95% acetonitrile, 5%water, 0.05% TFA; 17 mL/min; 0-50% B in 10 min., 50% B for 17 min.,50-100% B in 20 min). Fractions containing the product were combined andlyophilized to provide 24 mg of compound 121.

5-[({7-[3-(7-{[(3-carboxy-4-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))-2-oxoimidzoidinyl]-(2-naphthyl)}sulfonyl)amino]-2-hydroxybenzoicacid (Compound 122)

To 8 mg (0.011 mmol) of compound 121 in 2 mL of saturated sodiumcarbonate and 2 mL of water was added 3 mg (0.010 mmol) of triphosgenedissolved in 0.5 mL of THF, dropwise. The addition was made over aperiod of 15 min. The reaction was judged incomplete by HPLC, so another4.5 mg (0.015 mmol) of triphosgene in 0.5 mL of THF was added as before.After 2 hours, the reaction was acidified with 6N HCl and a precipitateformed. The suspension was frozen and lyophilized. The resulting solidwas dissolved in dimethyl sulfoxide/water/acetonitrile and purified byreverse-phase HPLC (C18, 250×20 mm column) using trifluoroacetic acid(TFA) buffer system (Buffer A: 5% acetonitrile, 95% water, 0.05% TFA;Buffer B: 95% acetonitrile, 5% water, 0.05% TFA; 19 mL/min; 0-100% B in20 min). This provided 1 mg of the desired compound 122.

Compound 123 was prepared by a similar synthetic sequence as outlined inReaction Scheme VI.

TABLE 6

Compound R 122 OH 123 Cl

Example 7

Compound 129 was prepared according to the procedure outlined inReaction Scheme VII. Compound 126 was prepared from commerciallyavailable 3-sulfobenzoic acid using standard procedures known to thoseskilled in the art.

naphthalene-2,7-diamine (Compound 125)

To 1.39 g (6.37 mmol) of 2,7-dinitronaphthalene (124) was added 25 mL ofconcentrated HCA and 15 mL of ethanol. Then, 9.6 g (50.9 mmol) of tin(II) chloride was added and the reaction was heated at 78 C for 24hours. The reaction was made basic with NaOH and extracted with ethylacetate. The ethyl acetate layer was dried (MgSO₄), filtered, and thevolatiles removed by rotary evaporation. The product was purified bysilica gel column chromatography eluting with 1% methanol indichloromethane. This provided 0.965 g of compound 125.

methyl 3-{[(7-amino-2-naphthyl)amino]sulfonyl}benzoate (Compound 127)

To 277 mg (1.19 mmol) of compound 125 was added 30 mL of THF. Then, 900μL of pyridine and 10 mL of sulfolane was added to this suspension. Asolution of 307 mg (1.31 mmol) of compound 126 in 10 mL of THF was addeddropwise. The reaction was allowed to stir at ambient temperature for 14hours. The reaction was extracted with ethyl acetate (2×) and 1N HCl.The ethyl acetate layers were discarded. The aqueous layer was madebasic with NaOH and then extracted with ethyl acetate. The ethyl acetatelayer was then extracted with water at pH 2.6 until no more startingmaterial (compound 125) was detected in the organic layer. The ethylacetate layer was then dried (MgSO₄), filtered, and the volatilesremoved by rotary evaporation. The resulting residue was dissolved inethyl acetate and treated with 1N HCl in diethyl ether. The resultingsolid was collected to yield 48 mg of compound 127 as the hydrochloridesalt.

methyl3-({[7-({[7-({[3-(methoxycarbonyl)phenyl]sulfonyl}amino)-2-naphthyl]amino}carbonylamino)-2-naphthyl]amino}sulfonyl)benzoate(compound 128) and3-{[(7-{[(7-{[(3-carboxyphenyl)sulfonyl]amino}-2-naphthyl)amino]carbonylamino}-2-naphthyl)amino]sulfonyl}benzoicacid (Compound 129)

To 43 mg (0.11 mmol) of compound 127 was added 8 mL of 1 M sodiumbicarbonate and 0.5 mL of THF to give a clear solution. Then, a solutionof 33 mg (0.11 mmol) of triphosgene in 0.5 mL of THF was added dropwise.HPLC analysis showed the reaction was incomplete so another 33 mg oftriphosgene was added dropwise. After HPLC analysis showed the reactionto still be incomplete, a third batch of triphosgene was added. Thereaction was judged complete so the volatiles were removed by rotaryevaporation and the resulting residue (compound 128) was treated with 2NNaOH. The reaction was allowed to stir at ambient temperature for 14hours. Then, the solution was adjusted to pH 1 with 6 N HCl, and aprecipitate formed. The solid was collected by vacuum filtration andpurified by silica gel column chromatography to provide 16 mg ofcompound 129.

Example 8

The unsymmetrical compounds 136-145 were prepared according to thegeneral procedures outlined in Reaction Scheme VIII for the synthesis ofcompounds 136 and 137. The various amines used in place of compound 18were prepared by the general procedure for compound 18 that is outlinedin Reaction Scheme II.

7-(acetylamino)naphthalene-2-sulfonic acid sodium salt (Compound 130)

To 3.75 g (16.8 mmol) of 7-amino-naphthalene-2-sulfonic acid(compound 1) was added 20 mL each of pyridine and acetic anhydride. Thereaction was allowed to stir at ambient temperature for 24 hours. Theblack reaction was cooled in an ice bath and then 45 mL of methanol wasadded slowly. After 1 hour, a solution of sodium methoxide (425 mg ofsodium in 10 mL methanol) was added. A precipitate formed. Thesuspension was allowed to stir for 2 hours and then the solid wascollected by vacuum filtration and was washed with ethyl acetate. Thisprovided 4.4 g of compound 130.

N-[7-(chlorosulfonyl)-2-naphthyl]acetamide (Compound 131)

To 3.6 g (12.5 mmol) of compound 130 was added 100 mL of phosphorousoxychloride. Then, 4 mL of dimethylacetamide was added dropwise. Thereaction was allowed to stir at ambient temperature for 5 hours and thenwas poured onto 2 L of ice. After the ice had melted, the solidprecipitate was collected by vacuum filtration and was washed withwater. After drying in vacuo, this provided 3.4 g of compound 131.

5-({[7-(acetylamino)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoic acid(Compound 132)

To 15 g (0.087 mol) of 5-amino-2-chlorobenzoic acid was dissolved in 450mL of THF and 15 mL of pyridine. The solution was cooled to 5° C. in anice-water bath. Then, a solution of 20.8 g (0.074 mol) of compound 131dissolved in 200 mL of THF was added over a 10 min period. The reactionwas kept at 5° C. for 30 min, and then allowed to warm to roomtemperature. The reaction was allowed to stir for an additional 4 hours.Then, the reaction was filtered to remove some insoluble material andthe resulting clear filtrate was reduced to a solid by the removal ofthe volatiles by rotary evaporation. This solid was extracted with ethylacetate and 1 N HCl. The ethyl acetate layer was further extracted with0.5 M NaOH (once) and 0.33 M NaOH (three times). The aqueous layers werecombined, acidified with HCl, and back extracted into ethyl acetate.After drying (MgSO₄), filtration, and removal of the ethyl acetate byrotary evaporation, the solid residue was dissolved in 2.8 L of 50/50methanol/water with heating. The solution was allowed to cool to roomtemperature and a small amount of solid was removed by vacuum filtrationand discarded. The clear filtrate was allowed to sit for 48 hours andthen was reduced in volume to about 500 mL by rotary evaporation. Thesolid was collected by vacuum filtration. After drying in vacuo, thisprovided 17.6 g of compound 132.

5-{[(7-amino(2-naphthyl))sulfonyl]amino}-2-chlorobenzoic acid (Compound133)

To 13.5 g (0.032 mol) of compound 132 was added 100 mL of 5N NaOH. Thissolution was heated at 50° C. for 8 hours. Then, the reaction wasacidified with 86 mL of 6N HCl and extracted with ethyl acetate. Theethyl acetate was washed with 1N HCl, water, and brine. The organiclayer was dried (MgSO₄), filtered, and the volatiles removed by rotaryevaporation to yield 11.3 g of compound 133.

methyl 5-{[(7-amino(2-naphthyl))sulfonyl]amino}-2-chlorobenzoate(Compound 134)

To 10.1 g (0.027 mol) of compound 133 dissolved in 250 mL of methanolwas added 50 mL of 4 N HCl in dioxane. This solution was allowed to stirat ambient temperature for 18 hours. The reaction was incomplete, so itwas heated at reflux for an additional 5 hours. Then, the volatiles wereremoved by rotary evaporation and the resulting solid was extracted withethyl acetate and 0.4 N sodium bicarbonate, water, and brine. Theorganic layer was dried (MgSO₄), filtered, and the volatiles removed byrotary evaporation to yield 8.71 g of compound 134.

methyl2-chloro-5-{[(7-isothiocyanato(2-naphthyl))sulfonyl]amino}benzoate(Compound 135)

To 4.5 g (11.5 mmol) of compound 134 and 9.01 g (50.6 mmol) of1,1′-thiocarbonyldiimidazole was added 50 mL of TBF. The solution wasallowed to stir at ambient temperature for 1.5 hours. Then, the reactionwas poured into 300 mL of ethyl acetate and extracted with 1N HCl,water, and brine. The organic layer was dried (MgSO₄), filtered, and thevolatiles removed by rotary evaporation to yield 4.77 g of compound 135.

3-[({7-[({[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino)-}-thioxomethyl)amino]-4-hydroxy-2-naphthyl}sulfonyl)amino]benzenesutfonicacid (Compound 136)

To 170 mg (0.39 mmol) of compound 135 dissolved in 5 mL of DMF was addeda solution of 100 mg (0.25 mmol) of compound 18 in 5 mL of DMF. Thereaction was allowed to stir at ambient temperature. After 4 days, theDMF was removed by rotary evaporation and kept under high vacuum toremove traces of DMF. Then, the resulting residue was treated withdichloromethane (50 mL) and sonicated to form a suspension. The solid,insoluble product was collected by vacuum filtration. This solid wasdissolved in methanol and then the methanol was removed by rotaryevaporation. The resulting solid was again treated with dichloromethane,sonicated, and collected by vacuum filtration. This provided 172 mg ofan orange solid.

2-chloro-5-{[(7-{[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]-carbonylamino}(2-naphthyl))sulfonyl]amino}benzoicacid (Compound 137)

To 100 mg (0.12 mmol) of compound 136 was added 15 mL of acetonitrilefollowed by 10 mL of THF and 1 mL of iodomethane. The reaction wasallowed to stir at ambient temperature for 4 days. HPLC analysisindicated that the reaction was complete. The volatiles were removed byrotary evaporation and the residue was dissolved in 1N NaOH (aqueous) at0-50 C. The reaction was kept in an ice bath for 30 minutes, and thenallowed to warm to room temperature over 2 hours. The reaction solutionwas filtered through a 0.2 um nylon filter to remove some cloudiness.Then the reaction pH was adjusted to about 1 with 6N HCl. The resultingsolid precipitate was collected by vacuum filtration. This provided 60mg of compound 137.

The compounds 138-145 (Table 7) were prepared by the same generalprocedures as described above for the synthesis of compounds 136 and 137except that different amines were used in place of compound 18. Theamines were prepared by the same general procedure as for the synthesiscompound 18, as depicted in Reaction Scheme II. Compounds in Table 7with are shown in the free acid form.

TABLE 7

Compound # R⁰ R⁵ R⁶ R⁰⁰ K 136

OH H

S 137

OH H

S 138

OH H

O 139

H H

S 140

H H

S 141

H H

O 142

H H

S 143

H H

S 144

H H

O 145

H H

S 146

H H

S 147

H H

O 148

OH H

O 149

OH H

S 150

OH H

O 151

H H

O 152

H H

O 153

H H

O 154

H H

S 155

H H

S 156

H H

O

Example 9

The compounds 157-168 were prepared according to the procedures outlinedin Reaction Scheme IX.

methyl2-chloro-5-[({7-[({[7-({[4chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}thioxomethyl)amino](2-naphthyl)}sulfonyl)amino]benzoate(Compound 157)

To 3.0 g (7.68 mmol) of compound 134 was added a solution of 4.0 g (9.24mmol) of compound 135 in 200 mL of dichloromethane. The reaction wasallowed to stir at ambient temperature for 48 hours. The fine, whitesolid was collected to give 5.2 g of compound 157.

methyl5-({[7-(1-aza-2-{[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino)-2-methylthiovinyl)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoate(Compound 158)

To 1.0 g (1.21 mmol) of compound 157 dissolved in 50 mL of acetonitrilewas added 1.1 mL of methyl iodide (17.7 mmol). The reaction was allowedto stir under an argon atmosphere for 72 hours. The volatiles wereremoved by rotary evaporation, and the resulting yellow solid wasextracted using ethyl acetate and 1M sodium carbonate. The organic layerwas washed with 50/50 brine/water followed by brine. The organic layerwas separated and volatiles removed by rotary evaporation to yield 0.95g of compound 158.

methyl5-({[7-(2-amino-1-aza-2-{[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}vinyl)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoate(Compound 159)

To 220 mg (0.24 mmol) of compound 158 was added 10 mL of a 0.5 M NH₃solution in dioxane. The resulting solution was placed in a sealed tubeand heated at 70° C. for 18 hours, followed by 24 hours at 80° C. Then,additional NH₃ gas was bubbled into the reaction for 4 min. The reactionwas sealed and heating at 80° C. continued for an additional 51 hours.Then, the reaction temperature was lowered to 65° C. and continued for11 days. At this point, the reaction had reached 70% completion basedupon HPLC analysis. The reaction was stopped by removal of the volatilesby rotary evaporation. The product was purified by silica gel columnchromatography eluting with 3% methanol in dichloromethane followed by5% methanol in dichloromethane. Finally, the product was eluted with90:2:1 ethyl acetate:isopropanol:water. This provided 117 mg of compound159.

5-{[(7-{2-amino-1-aza-2-[(7-{[(3-carboxy-4-chlorophenyl)amino]sulfonyl}(2-naphthyl)amino]vinyl}(2-naphthyl))sulfonyl]amino}-2-chlorobenzoicacid (Compound 160)

To 50 mg (0.062 mmol) of compound 159 was added 10 mL of 1N NaOH. Theresulting solution was allowed to stir at ambient temperature for 1hour. The reaction was acidified to pH 1 using 11 mL of 1N HCl. A whiteprecipitate formed, which was collected by vacuum filtration and waswashed with water. The solid was dried in vacuo to provide 45 mg ofcompound 160.

The compounds 161-168 (Table 8) were prepared by the same. generalprocedures as described above for the synthesis of compounds 157-160except that other amines were used in place of ammonia. Compounds inTable 8 with acidic groups are shown in the free acid form.

TABLE 8

Compound # K R 157 S OCH₃ 158 SCH₃ OCH₃ 159 NH OCH₃ 160 NH OH 161 NCH₃OCH₃ 162 NCH₃ OH 163 N(CH₃)₂ OCH₃ 164 N(CH₃)₂ OH 165 NCN OCH₃ 166 NCN OH167 NHCH₂CH═CH₂ OCH₃ 168 NHCH₂CH═CH₂ OH

Example 10

The compounds 169-181 (Table 9) were prepared according to the samegeneral procedures outlined in Reaction Schemes I and II and describedfor compounds in Table 1. The compound 6-naphthalene-2-sulfonic acid wasused in place of compounds 1 and 2.

TABLE 9

Compound # R⁰ R⁰⁰ 169

170

171

172

173

174

175

176

177

178

179

180

181

Example 11

The [¹⁴C] labeled compound 15 was prepared according to the procedureoutlined in Reaction Scheme X.

[¹⁴C]-3-{[(4-hydroxy-7-{[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]-carbonylamino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid (Compound [¹⁴C]-15)

To 51 mg (0.013 mmol) of compound 18 dissolved in 1.7 mL of DMF wasadded 58 μL (0.026 mmol) of 2,6-di-tert-butylpyridine. Then, in aseparate test tube with 13.6 mg (0.020 mmol) of imidazole in 1.7 mL ofTHF, was added 36 μL (0.007 mmol) of [¹⁴C]-phosgene (20% in toluene).This formed a white solid. After 2 min., 1.7 mL of DMF was added to theTHF suspension to form a clear solution. This solution was added to theDMF solution of compound 18. After 4 hours, the product was purified byreverse-phase HPLC (C18, 250×20 mm column) using trifluoroacetic acid(TFA) buffer system (Buffer A: 5% acetonitrile, 95% water, 0.1% TFA;Buffer B: 95% acetonitrile, 5% water, 0.1% TFA) to give 16 mg ofcompound [¹⁴C]-15 with an activity of 55 mCi/mmol. names of thecompounds prepared according to the general procedures described andwhose structures are indicated in Tables 1-9 are listed in Table 10.Compounds that have acidic functionalities are named as the parent freeacid. The following IUPAC names were derived using the software programChemistry 4D Draw™ from ChemInnovation Software, Inc.

TABLE 10 Compound # IUPAC Name 37-{[(7-sulfo-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic acid44-hydroxy-7-{[(5-hydroxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid 87-[({7-[({3-[(4-methylphenyl)oxysulfonyl)phenyl}amino)sulfonyl]-2-naphthyl}amino)carbonylamino]naphthalene-2-sulfonic acid 94-methylphenyl3-[({7-[(N-{7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate 104-hydroxy-7-[({5-hydroxy-7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl](2-naphthyl)}amino)carbonylamino]naphthalene-2-sulfonic acid 114-methylphenyl 3-[({4-hydroxy-7-[(N-{5-hydroxy-7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl](2-naphthyl)}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate 127-{[(7-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic acid 133-{[(7-{[N-(7-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulfonic acid 144-hydroxy-7-{[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonic acid 153-{[(4-hydroxy-7-{[N-(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid 163-bromo-7-{[(6-bromo-5-hydroxy-7-sulfo(2-naphthyl))amino]carbonylamino}-4-hydroxynaphthalene-2-sulfonic acid 197-[({7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}amino)carbonylamino]naphthalene-2-sulfonic acid 207-{[(7-{[(4-sulfophenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic acid 21 4-methylphenyl4-[({7-[(N-{7-[({4-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]benzenesulfonate 224-{[(7-{[N-(7-{[(4-sulfophenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulfonic acid 23N-(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide 24 methyl4-({[7-({[7-({[4-methoxycarbonyl)phenyl]amino}sulfonyl-2-naphthyl]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 254-{[(7-{[N-(7-{[(4-carboxyphenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzoic acid 264-hydroxy-7-({[5-hydroxy-7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)naphthalene-2-sulfonic acid 27 methyl4-({[4-hydroxy-7-({[5-hydroxy-7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 284-{[(4-hydroxy-7-{[N-(5-hydroxy-7-sulfo(2-naphthyl))carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzoic acid 294-{[(7-{[N-(7-{[(4-carboxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}4-hydroxy-2-naphthyl)sulfonyl]amino}benzoic acid 302-hydroxy-3-{[(7-{[N-(7-sulfo(2-naphthyl)}carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 313-{[(7-{[N-(7-{[(3-carboxy-2-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoic acid 322-hydroxy-5-{[(7-{[N-(7-sulfo(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 335-{[(7-{[N-(7-{[(3-carboxy-4-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoic acid 342-hydroxy-4-{[(7-{[N-(7-sulfo(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 354-{[(7-{[N-(7-{[(4-carboxy-3-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoic acid 367-({[7-({[3-(methoxycarbonyl)-4-nitrophenyl]amino}sulfonyl)-2-naphthyl]amino}carbonylamino)naphthalene-2-sulfonic acid 372-nitro-5-{[(7-{[N-(7-sulfo(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoicacid 385-{[(7-{[N-(7-{[(3-carboxy-4-nitrophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-nitrobenzoic acid 39 methyl3-({[7-({[7-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl)-2-naphthyl]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 403-{[(7-{[N-(7-{[(3-carboxyphenyl)amino]sulfonyl}-2-naphthyl}carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzoic acid 41 methyl3-({[4-hydroxy-7-({[5-hydroxy-7-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 423-{[(7-{[N-(7-{[(3-carboxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}4-hydroxy-2-naphthyl)sulfonyl]amino}benzoic acid 435-{[(7-{[N-(7-{[(3-carboxy-4-hydroxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}-4-hydroxy(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoicacid 44 methyl2-({[7-({[7-({[2-(methoxycarbonyl)phenyl]amino}sulfonyl)-2-naphthyl]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 452-{[(7-{[N-(7-{[(2-carboxyphenyl)amino]sulfonyl}-2-naphthyl)carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzoic acid 46 methyl2-({[4-hydroxy-7-({[5-hydroxy-7-({[2-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 472-{[(7-{[N-(7-{[(2-carboxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}4-hydroxy-2-naphthyl)sulfonyl]amino}benzoic acid 485-{[(7-{[N-(7-{[(3-carboxy-4-chlorophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-chlorobenzoic acid 495-{[(7-{[N-(7-{[(3-carboxy-4-chlorophenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}-4-hydroxy(2-naphthyl))sulfonyl]amino}-2-chlorobenzoicacid 50 methyl3-({[7-({[7-({[3,5-bis(methoxycarbonyl)phenyl]amino}sulfonyl)-5-hydroxy(2-naphthyl)]amino}carbonylamino)-4-hydroxy(2-naphthyl)]sulfonyl}amino)-5-(methoxycarbonyl)benzoate 515-{[(7-{[N-(7-{[(3,5-dicarboxyphenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}4-hydroxy-2-naphthyl)sulfonyl]amino}benzene-1,3-dicarboxylicacid 522-[3-({[7-({[7-({[3-(carboxymethyl)phenyl]amino}sulfonyl)-5-hydroxy(2-naphthyl)]amino}carbonylamino)-4-hydroxy-2-naphthyl]sulfonyl}amino)phenyl]aceticacid 53 methyl2-hydroxy-5-({[7-({[7-({[4-hydroxy-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}amino)benzoate 54methyl2-chloro-5-({[7-({[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}amino)benzoate 55methyl2-bromo-5-({[7-({[7-({[4-bromo-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}amino)benzoate 562-bromo-5-{[(7-{[N-(7-{[(4-bromo-3-carboxyphenyl)amino]sulfonyl}(2-naphthyl)}carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 575-{[(7-{[N-(7-{[(3-carboxy-4-methylphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-methylbenzoic acid 585-{[(7-{[N-(7-{[(3-carboxy-4-fluorophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-fluorobenzoic acid 593-{[(7-{[N-(7-{[(3-carboxy-2-chlorophenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}-4-hydroxy(2-naphthyl))sulfonyl]amino}-2-chlorobenzoicacid 603-{[(7-{[N-(7-{[(3-carboxy-6-chlorophenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}-4-hydroxy(2-naphthyl))sulfonyl]amino}-4-chlorobenzoicacid 61 methyl2-methoxy-5-({[7-({[7-({[4-methoxy-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}amino)benzoate 625-{[(7-{[N-(7-{[(3-carboxy-4-methoxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-methoxybenzoic acid 63 methyl4-hydroxy-3-({[7-({[7-({[2-hydroxy-5-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}amino)benzoate 643-{[(7-{[N-(7-{[(3-carboxy-6-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}4-hydroxybenzoic acid 65 methyl4-methoxy-3-({[7-({[7-({[2-methoxy-5-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}amino)benzoate 663-{[(7-{[(7-{[(3-carboxy-6-methoxyphenyl)amino]sulfonyl}(2-naphthyl))amino]carbonylamino}(2-naphthyl))sulfonyl]amino}-4-methoxybenzoicacid 67N-(7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide 68N-(5-hydroxy-7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))[(5-hydroxy-7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide69N-(5-hydroxy-7-{[(4-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))[(5-hydroxy-7-{[(4-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide70N-[7-({[3-(diethoxyphosphoryl)phenyl]amino}sulfonyl)(2-naphthyl)]{[7-({[3-(diethoxyphosphoryl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide71N-[7-({[3-(ethoxy(hydroxyphosphoryl))phenyl]amino}sulfonyl)(2-naphthyl)]{[7-({[3-(ethoxy(hydroxyphosphoryl))phenyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide727-({[7-({[(1S)-2-(4-hydroxyphenyl)-1-(methoxycarbonyl)ethyl]amino}sulfonyl)-2-naphthyl]amino}carbonylamino)naphthalene-2-sulfonic acid 73(2S)-3-(4-hydroxyphenyl)-2-{[(7-{[N-(7-sulfo(2-naphthyl)}carbamoyl]amino}(2-naphthyl))sulfonyl]amino}propanoic acid 74 methyl(2S)-2-({[7-({N-[7-({[(1S)-2-(4-hydroxyphenyl)-1-(methoxycarbonyl)ethyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)-3-(4-hydroxyphenyl)propanoate 75(2S)-2-({[7-({N-[7-({[(1S)-1-carboxy-2-(4-hydroxyphenyl)ethyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)-3-(4-hydroxyphenyl)propanoicacid 76N-{7-[(phenylamino)sulfonyl](2-naphthyl)}({7-[(phenylamino)sulfonyl](2-naphthyl)}amino)carboxamide 77N-{7-[(3-pyridylamino)sulfonyl](2-naphthyl)}({7-[(3-pyridylamino)sulfonyl](2-naphthyl)}amino)carboxamide 787-[({7-[(pyrimidin-2-ylamino)sulfonyl]-2-naphthyl}amino)carbonylamino]naphthalene-2-sulfonicacid 79N-{7-[(pyrimidin-2-ylamino)sulfonyl](2-naphthyl)}({7-[(pyrimidin-2-ylamino)sulfonyl](2-naphthyl)}amino)carboxamide 80N-{7-[(pyrazin-2-ylamino)sulfonyl](2-naphthyl)}({7-[(pyrazin-2-ylamino)sulfonyl](2-naphthyl))amino)carboxamide 81N-(7-{[(3-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide 82N-[5-hydroxy-7-({[3-(hydroxymethyl)phenyl]amino}sulfonyl)(2-naphthyl)]{[5-hydroxy-7-({[3-(hydroxymethyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide 83N-(7-{[(3-cyanophenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-cyanophenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide 84N-(7-{[(3-nitrophenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-nitrophenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide 85N-[7-({[4-chloro-3-(hydroxymethyl)phenyl]amino}sulfonyl)(2-naphthyl)]{[7-({[4-chloro-3-(hydroxymethyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide 86N-(7-{[(3-chloro-4-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))[(7-{[(3-chloro-4-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide 87N-[7-({[(3,4-dihydroxyphenyl)methyl]amino}sulfonyl)(2-naphthyl)]{[7-({[(3,4-dihydroxyphenyl)methyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide 88N-[5-hydroxy-7-({[3-(trifluoromethyl)phenyl]amino}sulfonyl)(2-naphthyl)]{[5-hydroxy-7-({[3-(trifluoromethyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide 89N-(7-sulfanoyl(2-naphthyl))[(7-sulfamoyl(2-naphthyl))amino]carboxamide907-{[(7-{[(4-chloro-3-sulfophenyl)amino]sulfonyl}(2-naphthyl)amino]carbonylamino}naphthalene-2-sulfonic acid 912-chloro-5-{[(7-{[(7-{[(4-chloro-3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]carbonylamino}(2-naphthyl))sulfonyl]amino}benzenesulfonicacid 927-{[(7-{[(4-chloro-3-sulfophenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))amino]carbonylamino}-4-hydroxynaphthalene-2-sulfonic acid 932-chloro-5-{[(7-{[(7-{[(4-chloro-3-sulfophenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))amino]carbonylamino}-4-hydroxy(2-naphthyl))sulfonyl]amino}benzenesulfonicacid 947-[({5-[(ethoxycarbonyl)methoxy]-7-sulfo(2-naphthyl)}amino)carbonylamino]4-hydroxynaphthalene-2-sulfonic acid 954-[(ethoxycarbonyl)methoxy]-7-[({5-[(ethoxycarbonyl)methoxy]-7-sulfo(2-naphthyl)}amino)carbonylamino]naphthalene-2-sulfonic acid 962-(6-{[N-(5-hydroxy-7-sulfo(2-naphthyl))carbamoyl]amino}-3-sulfonaphthyloxy]aceticacid 972-[6-({N-[5-(carboxymethoxy)-7-sulfo(2-naphthyl)]carbamoyl}amino)-3-sulfonaphthyloxy]aceticacid 984-hydroxy-7-{[(5-methoxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid 994-methoxy-7-{[(5-methoxy-7-sulfo(2-naphthyl))amino]carbonylamino}naphthalene-2-sulfonicacid 1004-[(2-sulfophenyl)methoxy]-7-[({7-sulfo-5-[(2-sulfophenyl)methoxy](2-naphthyl)}amino)carbonylamino]naphthalene-2-sulfonic acid 1014-(3-sulfopropoxy)-7-({[7-sulfo-5-(3-sulfopropoxy)(2-naphthyl)]amino}carbonylamino)naphthalene-2-sulfonic acid 1024-hydroxy-7-({[7-sulfo-5-(3-sulfopropoxy)(2-naphthyl)]amino}carbonylamino)naphthalene-2-sulfonic acid 1033-{methyl[(7-{[(7-{[methyl(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]carbonylamino}(2-naphthyl))sulfonyl]amino}benzenesulfonicacid 104 prop-2-enyl2-chloro-5-({[7-({[7-({[4-chloro-3-(prop-2-enyloxycarbonyl)phenyl]prop-2-enylamino}sulfonyl)(2-naphthyl)]amino}carbonylamino)(2-naphthyl)]sulfonyl}prop-2-enylamino)benzoate 1055-{[(7-{[(7-{[(3-carboxy-4-chlorophenyl)prop-2-enylamino]sulfonyl}(2-naphthyl))amino]carbonylamino}(2-naphthyl))sulfonyl]prop-2-enylamino}-2-chlorobenzoicacid 112 4-methylphenyl3-{[7-({[7-(N-{3-[(4-methylphenyl)oxysulfonyl]phenyl}carbamoyl)-2-naphthyl]amino}carbonylamino)-2-naphthyl]carbonylamino}benzenesulfonate113 methyl 3-[(7-{[(7-{N-[3-(methoxycarbonyl)phenyl]carbamoyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)carbonylamino]benzoate 1143-({7-[({7-[N-(3-sulfophenyl)carbamoyl]-2-naphthyl}amino)carbonylamino]-2-naphthyl}carbonylamino)benzenesulfonic acid 1163-({7-[({7-[N-(3-carboxyphenyl)carbamoyl]-2-naphthyl}amino)carbonylamino]-2-naphthyl}carbonylamino)benzoic acid 1225-[({7-[3-(7-{[(3-carboxy-4-hydroxyphenyl)amino]sulfonyl}(2-naphthyl))-2-oxoimidazolidinyl](2-naphthyl)}sulfonyl)amino]-2-hydroxybenzoic acid 1235-[({7-[3-(7-{[(3-carboxy-4-chlorophenyl)amino]sulfonyl}(2-naphthyl))-2-oxoimidazolidinyl](2-naphthyl)}sulfonyl)amino]-2-chlorobenzoic acid 1293-{[(7-{[N-(7-{[(3-carboxyphenyl)sulfonyl]amino}-2-naphthyl]carbamoyl]amino)-2naphthyl)amino]sulfonyl}benzoic acid 1363-[({7-[({[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}thioxomethyl)amino]-4-hydroxy-2-naphthyl}sulfonyl)amino]benzenesulfonicacid 1372-chloro-5-({[7-({[(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)benzoicacid 1382-chloro-5-{[(7-{[N-(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 139methyl2-chloro-5-[({7-[({[7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}thioxomethyl)amino](2-naphthyl)}sulfonyl)amino]benzoate1405-({[7-({[(7-{[(4-carboxyphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoic acid 1415-{[(7-{[N-(7-{[(4-carboxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-chlorobenzoic acid 142 methyl2-chloro-5-[({7-[({[7-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}thioxomethyl)amino](2-naphthyl)}sulfonyl)amino]benzoate1435-({[7-({[(7-{[(3-carboxyphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoic acid 1445-{[(7-{[7-{[(3-carboxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino)(2-naphthyl))sulfonyl]amino}-2-chlorobenzoic acid 145 methyl2-chloro-5-({[7-({[(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)benzoate146 2-chloro-5-({[7-({[(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)benzoicacid 1472-chloro-5-{[(7-{[N-(7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 148 methyl2-chloro-5-({[7-({[(5-hydroxy-7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)benzoate1492-chloro-5-({[7-({[(5-hydroxy-7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)benzoicacid 150 2-chloro5-{[(7-{[N-(5-hydroxy-7-{[(3-sulfamoylphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 151methyl 2-chloro-5-{[(7-{[({7-[(3-pyridylamino)sulfonyl](2-naphthyl)}amino)thioxomethyl]amino}(2-naphthyl))sulfonyl]amino}benzoate1522-chloro-5-{[(7-{[({7-[(3-pyridylamino)sulfonyl](2-naphthyl)}amino)thioxomethyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 1532-chloro-5-[({7-[N-{7-[(3-pyridylamino)sulfonyl](2-naphthyl)}carbamoyl)amino](2-naphthyl)}sulfonyl)amino]benzoic acid 154 methyl2-chloro-5-({[7-({[(7-{[(3-methylphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl)]sulfonyl}amino)benzoate155 2-chloro-5-({[7-({[(7-{[(3-methylphenyl)amino]sulfonyl}(2-naphthyl))amino]thioxomethyl}amino)(2-naphthyl))sulfonyl}amino)benzoicacid 1562-chloro-5-{[(7-{[N-(7-{[(3-methylphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}benzoic acid 157 methyl2-chloro-5-[({7-[({[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}thioxomethyl)amino](2-naphthyl)}sulfonyl)amino]benzoate158 methyl5-({[7-((1Z)-1-aza-2-{[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}-2-methylthiovinyl)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoate159 methyl2-chloro-5-[({7-[({[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl))amino}iminomethyl)amino](2-naphthyl)}sulfonyl)amino]benzoate160 5-({[7-({[(7-{[(3-carboxy-4-chlorophenyl)amino]sulfonyl}(2-naphthyl))amino]iminomethyl}amino)(2-naphthyl)]sulfonyl}amino)-2-chlorobenzoicacid 161 methyl5-[({7-[((1E)-2-aza-1-{[7-({[4-chloro-3-methoxycarbonyl)phenyl]sulfonyl}amino)(2-naphthyl)]amino}prop-1-enyl)amino](2-naphthyl)}amino)sulfonyl]-2-chlorobenzoate1625-({[7-({(1E)-2-aza-1-[(7-{[(3-carboxy-4-chlorophenyl)sulfonyl]amino}(2-naphthyl))amino]prop1-enyl}amino)(2-naphthyl)]amino}sulfonyl)-2-chlorobenzoic acid 163methyl 5-({[7-({(1Z)-2-aza-1-dimethylamino)-2-[7-({[4-chloro-3-(methoxycarbonyl)phenyl]sulfonyl}amino)(2-naphthyl)]vinyl)amino)(2-naphthyl)]amino}sulfonyl)-2-chlorobenzoate 1645-{[(7-{(1Z)-1-aza-2-(dimethylamino)-2-[(7-{[(3-carboxy-4-chlorophenyl)sulfonyl]amino)(2-naphthyl))amino]vinyl}(2-naphthyl))amino]sulfonyl}-2-chlorobenzoic acid165 methyl5-[((7-[((1E)-2-aza-1-{[4-chloro-3-methoxycarbonyl)phenyl]sulfonyl}amino)(2-naphthyl)]amino}-2-cyanovinyl)amino](2-naphthyl)}amino)sulfonyl]-2-chlorobenzoate1665-({[7-({(1E)-2-aza-1-[(7-{[(3-carboxy-4-chlorophenyl)sulfonyl]amino}(2-naphthyl))amino]-2-cyanovinyl}amino)(2-naphthyl)]amino}sulfonyl)-2-chlorobenzoic acid 167methyl5-[({7-[((1E)-2-aza-1-([7-({[4-chloro-3-methoxycarbonyl)phenyl]sulfonyl}amino)(2-naphthyl)]amino}penta-1,4-dienyl)amino](2-naphthyl)}amino)sulfonyl]-2-chlorobenzoate1685-({[7-({(1E)-2-aza-1-[(7-{[(3-carboxy-4-chlorophenyl)sulfonyl]amino}(2-naphthyl))amino]penta-1,4-dienyl}amino)(2-naphthyl)]amino}sulfonyl)-2-chlorobenzoicacid 1693-{[(6-{[(6-{[(3-sulfophenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)sulfonyl]amino}benzenesulfonic acid 170 methyl3-({[6{[6-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl-2-naphthyl]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 1713-{[(6-{[(6-{[(3-carboxyphenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino)-2-naphthyl)sulfonyl]amino}benzoic acid 172 methyl 4-({[6-({[6-({[4methoxycarbonyl)phenyl]amino}sulfonyl)-2-naphthyl]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate 1734-{[(6-{[(6-{[(4-carboxyphenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)sulfonyl]amino}benzoic acid 174 methyl3-({[6-({N-[6-({[3,5-bis(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)-5-(methoxycarbonyl)benzoate1755-{[(6-{[(6-{[(3,5-dicarboxyphenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)sulfonyl]amino}benzene-1,3 dicarboxylic acid 176 methyl2,4,5-trifluoro-3({[6-({N-[6-({[2,5,6-trifluoro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)benzoate 1773-{[(6-{[(6-{[(3-carboxy-2,5,6-trifluorophenyl)amino]sulfonyl}(2-naphthyl))amino]carbonylamino}(2-naphthyl))sulfonyl]amino}-2,4,8-trifluorobenzoicacid 178 ethyl2-{4-[({6-[(N-{6-[({4-[(ethoxycarbonyl)methyl]phenyl}amino)sulfonyl]-2-naphthyl}carbamoyl)amino]-2-naphthyl}sulfonyl)amino]phenyl}acetate 1792-[4-({[6-({N-[6-({[4-(carboxymethyl)phenyl]amino}sulfonyl)-2-naphthyl]carbamoyl}amino)-2-naphthyl]sulfonyl)amino)phenyl]acetic acid 180 ethyl1-[(6-{[(6-{[3-ethoxycarbonyl)piperidyl]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)sulfonyl]piperidine-3-carboxylate (mixture of 2 enantiomers andone meso) 1811-({6-[({6-[(3-carboxypiperidyl)sulfonyl]-2-naphthyl}amino)carbonylamino]-2-naphthyl}sulfonyl)piperidine-3-carboxylic acid (mixture of 2 enantiomersand one meso)

Example 11

³²P-cytoplasmic Kinase Domain (CKD) Autophosphorylation Assay

The insulin signaling pathway is activated through stimulation of theinsulin receptor. A major component of this activation is thephosphorylation of a specific portion of the receptor called theβ-kinase domain. The complete β-kinase domain of the human insulinreceptor (CKD) was expressed in, and purified from, baculovirus. CKD(4.0 μg/ml), in a solution of 29 mM HEPES (pH 7.6), 0.05% Triton X-100,10 mM MgCl₂, 2 mM MnCl₂ (50 μl final volume), was combined with 50 μMATP, and 5 μCi ³²P-ATP (3000 Ci/mmol.). A test compound, or the vehicle(dimethyl sulfoxide (DMSO)) was added to a final DMSO concentration of1%. The mixture was incubated for 10 minutes at room temperature. Thereaction was terminated by the addition of 10 μl of 200 mM EDTA. A 30 μlvolume was removed, mixed with 5 μl of 6×Laemmeli sodium dodecyl sulfate(SDS) treatment buffer, and heated to 94° C. for 5 minutes. A 20 μlaliquot was then run on an SDS-PAGE gel. The radioactivity incorporatedinto the CKD band was quantified by phosphorimaging of the gel, orscintillation counting of the excised bands. The potency of a compoundfor increasing phosphorylation was expressed as % of the vehicle level.The results for this are shown in Table 11, below.

TABLE 11 Compound % Activity vs. Control 3 88 4 152 9 118 11 96 12 13113 135 14 110 15 125 16 144 23 80 26 124 27 112 28 124 29 106 39 93 40130 41 109 42 124 76 89 77 171 89 104 94 279 95 101 97 99 98 166 99 110100 79 101 104 102 115

Example 12

Whole-cell Phosphorylation Assay

The initial step in insulin signaling is phosphorylation of the insulinreceptor in response to insulin binding. NIH 3T3 cells overexpressinghuman insulin receptor (3T3HIR) were grown for 2 days at a density of2×10⁵/ml cells in 6 well culture dishes in DMEM with 10% FBS andL-glutamine. Prior to the experiment the cells were serum-starvedovernight with DMEM with 0.1% BSA. The following morning, the cells werewashed with PBS and the medium was replaced with 150 mM NaCl, 1.7 mMKCl, 0.9 mM CaCl₂, K₂HPO₄ (pH 7.4), to which were added either theexperimental compounds, or their vehicle (DMSO). Insulin or its vehicle(0.01% BSA) was diluted in the assay buffer (containing test compound orvehicle, respectively) to a final concentration of 2.5 nM. Afterincubating for 15 min, the cells were washed with cold PBS twice andlysed in 50 mM Tris.HCl, pH 7.4 150 mM NaCl, 0.25% sodium deoxycliolate,1% NP-40, 1 mM EGTA, 1 mM PMSF, 1 mM Na₃VO₄, 1 mM NaF, 1 ug/ml each ofAprotinin, Leupeptin and Pepstatin. The cell lysates were clarified bycentrifugation at 12000 rpm, and the supernatants were estimated forprotein concentration. Total cell lysate (˜20×g) was boiled with2×SDS-PAGE sample buffer for 3 min and loaded into 7.5% SDS-PAGE alongwith Amersham rainbow marker protein as a molecular weight standard.

After completing SDS-PAGE, the proteins were transferred ontoImmobilon-P membrane and Western analysis was carried out by incubatingthe blot with anti-phosphotyrosine antibody and developed by EnhancedChemiluminiscence (ECL), as Shown in FIG. 1. Shown in FIG. 2 is theincrease of autophosphorylation by Compounds 13 and 15 at variousconcentrations.

Example 13

Glucose Transport Activity Assay

Stimulation of the insulin receptor leads to the transport of glucosefrom the blood into cells, thus modulating blood glucose levels. 3T3 L1fibroblasts (ATCC) were grown in Dulbecco's modified Eagle's medium(DMEM) with 10% fetal bovine serum (FBS). The cells were plated at adensity of 3×10⁴ cells/well in 24-well plates. Two days after confluencewas reached, the cells were treated for 3 days with 0.5 mMisobutylmethylxanthine (IBMX), 1 μM dexamethasone, and 1.7 μM insulin.The cells were then transferred to DMEM with 10% FBS and supplementedwith 1.7 μM insulin for 2 more days. The cells were maintained in DMEMwith 10% FBS for an additional 4 days. Finally the cells weresertun-starved overnight in 0.1% bovine serum albumin (BSA) in DMEM.

The following day, the medium was replaced with 150 mM NaCl, 1.7 mM KCl,0.9 mM CaCl₂, K₂HPO₄ (pH 7.4) to which were added either theexperimental compounds or their vehicle (DMSO). Insulin or its vehicle(0.01% BSA) was diluted in the assay buffer (containing test compound orvehicle, respectively) to final concentration of 5.6 nM. Afterincubation for 30 min at 37° C., 5 μCi/ml ¹⁴C-2-deoxy-D-glucose wasadded, and the incubation was continued for additional 30 min at 37° C.The cells were then washed 3 times with ice-cold PBS/20 mM glucose andlysed in 250 μl of lysis buffer (50 mM HEPES pH 7.6, 1% Triton X-100)for 30 min at room temperature. Radioactivity in the lysate wasquantified by scintillation counting.

Once ¹⁴C-2-deoxy-D-glucose is transported into the cell, it is notreleased. Glucose transport is, therefore, proportional to the amount ofradioactivity in the lysate. The concentration of compound necessary toproduce an increase in glucose transport to greater than 150% of thevehicle control (generally representing the sum of the standarddeviation of the vehicle control plus the largest standard deviation ofa test sample) was recorded as the EC (effective concentration). Theresults are shown in Table 12. Shown in FIG. 3 is glucose uptake bycells at various concentrations when treated with Compound 13 andCompound 15.

TABLE 12 Concentration (μM) to reach Compound >150% activity of vehiclecontrol 3 3.2 4 32 13 3.2 15 3.2 16 32 20 3.2 22 3.2 23 10 25 3.2 26 3228 32 29 3.2 30 3.2 31 3.2 32 32 33 3.2 34 >32 35 32 37 3.2 38 3.2 39 3240 3.2 41 32 42 3.2 43 10 45 3.2 47 3.2 48 3.2 49 3.2 50 3.2 51 >32 523.2 56 3.2 57 3.2 59 32 60 >32 62 10 64 3.2 65 >32 66 3.2 67 3.2 68 3.269 >32 71 3.2 72 3.2 73 10 74 3.2 75 3.2 77 3.2 78 10 79 10 80 3.2 813.2 82 3.2 83 3.2 84 3.2 85 32 88 >32 89 3.2 90 3.2 91 3.2 93 3.2 94 3298 32 99 32 103 3.2 105 >32 114 3.2 116 >32 122 32 129 32 157 >32158 >32 159 3.2 160 3.2 161 >32 162 >32 165 3.2 166 3.2 167 >32 168 >32169 3.2 171 3.2 173 3.2 175 3.2 177 >32 179 >32 181 3.2

Example 14

Effect of Wortmannin and Cytochalasin on Glucose Uptake

The glucose pathway stimulated by insulin involves activation of P13Kinase. Wortmannin is a selective inhibitor of P13 Kinase and inhibitsinsulin-stimulated glucose transport 3T3 L1 adipocytes were pretreatedwith 100 nM Wortmannin and stimulated with the compounds in the presenceor absence of insulin. Wortmannin inhibited the stimulation of glucosetransport, measured as in Example 11, by either Compound 15 alone orCompound 15 plus 5.6 nM insulin. Insulin-stimulated glucose transport ismediated by glucose transporter proteins. Cytochalasin B is an inhibitorof glucose transporters and inhibits insulin-stimulated glucose uptake.Like Wortmannin, Cytochalasin B (10 μM) also inhibited the stimulationof glucose transport, measured as in Example 13, by either Compound 15alone or Compound 15 plus 5.5 Nm INSULIN. These results suggest that theactivation of glucose transport by the compounds utilizes the insulinsignaling machinery of the cells. Results are shown in FIGS. 4 and 5.

Example 15

Immunofluorescence Analysis of GLUT4 Mobilization in 3T3 L1 Adipocytes

Insulin-dependent transport of glucose into cells utilizes glucosetransporter proteins, such as GLUT4. Stimulation with insulin causesthese transporters to translocate from storage sites within the cellmembrane, where they facilitate glucose entry.

3T3 L1 adipocytes were grown and differentiated as in Example 11, exceptthat they were grown on a microscope chamber slide. The cells wereserum-starved with 0.1% BSA in DMEM for 16 hrs and stimulated with 56 μMCompound 15 alone, 100 nM Insulin for 1 hr at 37° C. The cells werefixed with 3.5% paraformaldehyde for 5 min and permeabilized with 0.2%saponin in 1% BSA, TBS for 5 min followed by incubation with anti-GLUT4antibody for 30 min at room temperature. The cells were extensivelywashed and incubated with FITC-conjugated secondary antibody for 30 minat room temperature. The cells were washed, air-dried and mounted withmounting medium and examined under confocal microscope (Stuart A. Rosset. al 1996, J. Biol. Chem. 271: 3328-3332.)

As shown in FIG. 6, in unstimulated cells, the GLUT4 immunofluorescencewas distributed throughout the cell (FIG. 6A). Following stimulation byinsulin, GLUT4 was seen primarily at the cell surface (FIG. 6B),consistent with insulin-induced translocation. Treatment of cells withCompound 15 also produced apparent translocation of GLUT4 to the cellsurface (FIG. 6C), consistent with its ability to stimulate glucosetransport into cells in an insulin-like manner.

Example 16

Selectivity vs. EGFR, PDGFR and IGFR

In order to determine the selectivity of these compounds for the insulinreceptor, their effects on other receptors that share a similarmechanism of activation with the insulin receptor were examined. Humanepidermoid carcinoma (A431) cells were plated at a density of 2×10⁵cells per well in 6-well dishes in DMEM with 10% FBS and L-Glutamine andallowed to grow to 75% confluence. Prior to the experiment, the cellswere serum-starved overnight with DMEM with 0.1% BSA. The followingmorning, the cells were washed with PBS and the medium was replaced with150 mM NaCl, 1.7 mM KCl, 0.9 mM CaCl₂, K₂HPO₄ (pH 7.4), to which wasadded either the experimental compound or its vehicle (DMSO). EpidermalGrowth Factor (EGF) or its vehicle (0.01% BSA) was diluted in the assaybuffer (containing test compound or vehicle respectively) to a finalconcentration of 2.5 ng/ml. After incubating for 15 min, the cells werewashed with cold PBS twice and lysed in 50 mM Tris.HCl pH 7.4, 150 mMNaCl, 0.25% sodium deoxycholate, 1% NP40, 1 mM EGTA, 1 mM PMSF, 1 mMNa₃VO₄, 1 mM NaF, 1 ug/ml each of Aprotinin, Leupeptin and Pepstatin.The cell lysates were clarified by centrifugation at 12000 rpm, and thesupernatants were estimated for protein concentration. Total cell lysate(˜20 μg) was boiled with 2×SDS-PAGE sample buffer for 3 min and loadedin a 7.5% SDS-PAGE along with Amersham rainbow marker protein as amolecular weight standard.

After completing SDS-PAGE, the proteins were transferred ontoImmobilon-P membrane and Western analysis was carried out by incubatingthe blot with anti-phosphotyrosine antibody and developed by EnhancedChemiluminiscence (ECL). Results are shown in FIG. 7.

Compound 15 did not increase the phosphorylation of EGFR in the presenceor absence of EGF. Using modifications of this protocol know to thoseskilled in the art, as well as the appropriate cell types, Compound 15was likewise found not to increase the phosphorylation of theinsulin-like growth factor type 1 (IGF-1) or the platelet-derived growthfactor (PDGF) receptors either in the absence or presence of theirendogenous ligands (1GF-1 and PDGF, respectively).

Example 17

Blood Glucose Level Determination in db/db Mouse

An accepted model for Type 2 diabetes which has been used to establishthe potential anti-diabetic activity of compounds is the db/db mouse.Seven to 9 week old male db/db mice (Jackson Laboratories, Bar Harbor,Me.), were used to the study of the effects of compounds on bloodglucose levels. Animals were kept in a 12 h/12 h light/dark cycle, andexperiments were initiated immediately after the dark period (7:00a.m.). Food was removed at this time and returned after the final bloodglucose measurement was taken.

Insulin (0.5 U/ml, Humulin R, Catalog HI-201, Lilly, Indianapolis, Ind.)was prepared by diluting 100 U/mL stock insulin 1:200 with PBS(phosphate buffered saline, Gibco, BRL). Compounds were prepared in avehicle of either PBS or 20% DMSO in PBS.

Five to 10 animals (average weight 40-50 g) were used in each treatmentcondition. The animals were injected subcutaneously with either 0.01 Uinsulin in PBS or PBS alone, followed by 0.1 mL of compound or itsvehicle delivered intraperitoneally. Blood samples were taken 0 min, 15min, 30 min, 1 hr, 2 hr and 4 hr after the administration of the drug orvehicle by tail bleeding. Glucose measurements were made with aGlucometer and Glucose strips (Bayer).

The resulting data are shown in FIGS. 8 and 9. In FIG. 8, blood glucoselevels at various time points are shown following injections withphosphate buffered saline (PBS) only, insulin in PBS, or with compound15 and insulin in PBS. The blood glucose levels are reported as thepercentage of the “0-time” values at the time points indicated. FIG. 9shows the effect of compound 15 alone (without added insulin) on bloodglucose levels in db/db mice. Blood glucose levels at various timepoints are shown in FIG. 9 following injection of db/db mice withcompound 15 together with its vehicle (DMSO) and insulin in PBS. Bloodglucose levels at various time points following injections either withPBS alone are also shown for comparison. From the data it can be seenthat compound 15 acts in an insulin-independent manner to lower bloodglucose levels.

Example 18

Lowering of Blood Glucose, Insulin and Triglycerides in the ob/ob MouseModel of Type 2 Diabetes

Another standard model of Type 2 diabetes is the ob/ob mouse. Male ob/obmice (C57 BL/6J-ob) were housed 3-5 to a cage with free access tostandard rodent chow pellets. After one week, they were given a singledose of Compound 15 (30 mg/kg, p.o.). Compound 15 produced a 13%decrease in blood glucose levels, a 42% decrease in plasma insulinlevels, and a 15% decrease in plasma triglyceride levels. Theconcomitant reduction of plasma glucose and insulin levels areconsistent with a reduction in insulin resistance. Results are shown inFIG. 10.

Example 19

Blood glucose Determination in the STZ/HFD Rat Model of Type 2 Diabetes

Representative compounds were also profiled in genetically normal ratsgiven a high-fat diet followed by treatment with a low dose ofstreptozotocin (STZ/HFD). This treatment regimen recreates both theinsulin resistance and the hyperglycemia seen in human Type 2 diabetes.Male CD rats (Jackson Labs, Bar Harbor Me.) were maintained according toNIH guidelines, housed two per cage, and fed either standard lab chow(Tekland Laboratory Diets, James Grain, San Jose, Calif.) or the samechow supplemented with chocolate bars, cookies, and potato chips suchthat their final diet contained 30% fat by weight (high-fat diet; HFD).After two weeks of this diet, the animals were given an injection offreshly-prepared streptozotocin (35 mg/kg, i.p.) and continued on theHFD. Animals that achieved glucose levels of 190 to 380 mg/dl were usedin this study. At the end of the 12-hour light/dark cycle, and justprior to the experiment, the animals were moved in cages with no foodavailable until 4 hours after treatment. The compound or vehicle (PBS)was given p.o. by gavage, and blood was sampled by approved IACUCprotocol using tail cap method. Glucose was determined using theGlucometer Elite (Bayer, Elkhart, Ind.).

Compound 15 produced a reduction in blood glucose levels beginning onehour after administration and persisting for the entire 6 hours of theexperiment (FIG. 11). The reduction was maximal (20%) 4 hours after theadministration of Compound 15. The potencies of other compounds in themodel shown in Table 13.

TABLE 13 Reduction in blood glucose levels in the STZ/HFD rat modelafter oral compound administration (30 mg/kg). Results are shown asmaximal reduction over the time at which it occurred. Compound Number %Reduction in Blood Glucose/Time (h) 41 20/4 13 20/1 29 18/6 48 24/2 9325/4

Example 20

Rat Muscle Phosphorylation

Muscle is a partcularly important tissue for taking up glucose from theblood in response to insulin receptor stimulation. Activation of muscleinsulin receptors is, therefore, an important factor in the control ofblood glucose levels. STZ/HFD rats prepared as in Example 18 were givenan oral dose of compound 15 (30 mg/kg), and muscle samples wereharvested at different time points (0.5 hr, 1 hr, 2 hr, 3 hr, 4 hr, 6hr) and homogenized in extraction buffer (50 mM Tris.HCl pH 7.4, 150 mMNaCl, 0.25% sodium deoxycholate, 1% NP40, 1 mM EGTA, 1 mM PMSF, 1 mMNa₃VO₄, 1 mM NaF, 1 ug/ml each of Aprotinin, Leupeptin and Pepstatin).The tissue homogenate was centrifuged at 12K for 30 min at 4° C., andthe supernatants were saved. An equal protein amount from each samplewas immunoprecipitated with anti-insulin receptor antibody for 2 hr at4° C., followed by another 1 hr incubation with Protein G-Agarose beads.The immunecomplexes were washed three times with the extraction buffer,and the samples were boiled in 2×SDS-PAGE loading buffer for 5 min at100° C. The samples were resolved in a 7.5% SDS-PAGE along with Amershamrainbow marker protein as a molecular weight standard.

After completing SDS-PAGE, the proteins were transferred ontoImmobilon-P membrane and Western analysis was carried out by incubatingthe blot with anti-phosphotyrosine antibody and developed by EnhancedChemiluminiscence (ECL). FIG. 12 shows that Compound 15 produced anincrease in phosphorylation of the insulin receptor with a time-courseconsistent with that of blood glucose reduction (FIG. 11).

Example 21

Multi-dosing in db/db Mice

Male db/db mice 7 to 8 weeks old (Jackson Laboratories, Bar Harbor,Maine) were given Compound 15 (56 mg/kg, p.o.) or an equivalent amountof its vehicle (PBS), as in Example 17, daily for 3 days. Blood sampleswere taken by an IACUC-approved protocol (tail cap method) immediatelybefore each dose or 3 hours later. Blood glucose levels were measuredusing a Glucometer Elite (Bayer, Elkhart, Ind.). Blood glucose levelsshowed little change two hours after PBS administration (FIG. 13). Incontrast, compound 15 lowered blood glucose levels 2 hours afteradministration on each of the 3 days by 14 to 29%.

Example 22

Multi-dosing in STZ/HFD Rats

STZ/HFD rats were prepared as in example 19. Compound 15 or anequivalent amount of the vehicle (PBS) was administered daily by theoral route at a dose of 30 mg/kg. Blood samples were taken by anIACUC-approved protocol (tail cap method) immediately before each doseand 4 and 6 (day one only) hours later. Blood glucose levels weremeasured using a Glucometer Elite (Bayer, Elkhart, Ind.). The bloodglucose levels of the vehicle-treated group fell over the course of theexperiment as the animals recovered from the low dose of STZ (FIG. 14).Compound 15 administration lowered blood glucose levels below those ofthe vehicle-treated group by 6 hours after administration, and thislowering was sustained throughout the remainder of the 3 days.

Example 23

Acute Toxicity

Compound 15 was administered in PBS vehicle to male db/db mice (i.p.)and male CD rats (p.o.) at 300 mg/kg; which is approximately 10 timesits effective dose. No acute toxicity was seen.

Example 24

Ames Test (Compound 4)

Compound 4 was tested for its ability to cause mutations in thehistidine operon of Salmonella typhimurium strains TA89, TA100, TA1535,and TA1537, and at the tryptophan operon of Escherichia coli strainWP2uvrA. These represent standard tests for the mutagenic potential ofcompounds and are collectively called the Ames Test. The compounds weretested at non-toxic doses (50 to 5000 μg/plate) in the absence ofexogenous activation and in the presence of induced rat liver S-9 pluscofactors. Under the conditions of this study, the compounds did notinduce any significant increase in the number of revertant colonies forany of the tester strains and were, therefore, judged negative in theSalmonella typhimurium/Escherichia coli Plate Incorporation MutationAssay.

Example 25

P450 Interactions

A major pathway for elimination of drugs from the body, which may limittheir effectiveness, is the cytochrome P450 system of the liver.Compound 15 did not inhibit the catalytic activity of the humancytochromes P450, CYPIA2, CYP2C9, CYP2CI9, CYP2D6, or CYP3A4. Inaddition, Compounds 15, 53, and 48 were not metabolized by rate livermicrosomes after a 1-hour incubation.

Example 26

Oral Pharmaceutical Composition Preparation-solid Dosage Formulation

A pharmaceutical composition for oral administration may be prepared bycombining the following:

% w/w Compound of this invention  10% Magnesium stearate 0.5% Starch2.0% HPM cellulose 1.0% Microcrystalline cellulose 86.5% 

The mixture may be compressed to tablets, or filled into hard gelatincapsules.

The tablet may be coated by applying a suspension of film former (e.g.,HPM cellulose), pigment (e.g., titanium dioxide) and plasticiser (e.g.,diethyl phthalate) and drying the film by evaporation of the solvent.The film coat can comprise 2.0% to 6.0% of the tablet weight, preferablyabout 3.0%.

Example 27

Oral Pharmaceutical Composition Preparation-capsule

A pharmaceutical composition of a compound of the invention suitable fororal administration may also be prepared by combining the following:

% w/w Compound of this invention 20% Polyethylene glycol 400 80%

The medicinal compound is dispersed or dissolved in the liquid carrier,with a thickening agent added, if required. The formulation is thenenclosed in a soft gelatin capsule by suitable technology.

Example 8

Pharmaceutical Composition for Parenteral Administration

A pharmaceutical composition for parenteral administration may beprepared by combining the following:

Preferred Level Compound of this invention 1.0% Saline 99.0% 

The solution is sterilized and sealed in sterile containers.

Example 28

Distribution of Compound-[14C]-15 After Oral Aministration

STZ/IFD rats, prepared as in Example 19, were given a single oral doseof 30 mg/kg compound-[14C]-15 labeled with 50 μCi of 14C prepared as inExample 11A. Two hours later the animals were euthanized and 200 mgsamples of various tissues were removed by an IACUC-approved protocol.The tissue samples were homogenized and their radioactivity content wasmeasured by scintillation counting. The highest levels of radioactivitywere found in the pancreas, liver, and thigh muscle. These correspondedto Compound 15 concentrations of approximately 780 nM, 200 nM, and 175nM, respectively. These concentrations would be sufficient to produceinsulin receptor phosphorylation in vitro. Lower levels of radioactivityindicative of 50 nM to 100 nM concentrations of the compound were foundin abdominal muscle, fat, kidney, and spleen. The amount ofradioactivity in the blood was not above background levels.

All documents cited in the above specification are herein incorporatedby reference. Various modifications and variations of the presentinvention will be apparent to those skilled in the art without departingfrom the scope and spirit of the invention. Although the invention hasbeen described in connection with specific preferred embodiments, itshould be understood that the invention as claimed should not be undulylimited to such specific embodiments. Indeed, various modifications ofthe described modes for carrying out the invention which are obvious tothose skilled in the art are intended to be within the scope of thefollowing claims.

We claim:
 1. A compound of the formula:

where R¹ and R² are substituents on the A rings and are, independently,—SO₂NR⁷ ₂, —C(O)NR⁷ ₂, —NR⁷SO₂R⁷, —NR⁷C(O)R⁷, —SO₂OR⁷, —C(O)OR⁷,—OSO₂R⁷, or —OC(O)R⁷, R³ and R⁴ are, independently, hydrogen or loweralkyl, or R³ and R⁴ together are —(CH₂)₂—, —(CH₂)₃—, or —(CH₂)₄—, R⁵ andR⁶ are, independently, hydrogen, alkyl, substituted alkyl, cyano, halo,nitro, —SR⁸, —C(O)R⁸, —SO₂OR⁸, —OSO₂R⁸, —SO₂NR⁸ ₂, —NR⁸SO₂R⁸, —OC(O)R⁸,—C(O)OR⁸, —C(O)NR⁸ ₂, —NR⁸C(O)R⁸, —OR⁸, or —NR⁸ ₂, each R⁷ and R⁸ is,independently, hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, aryl(lower)alkyl, substituted aryl(lower)alkyl,heteroaryl(lower)alkyl, substituted heteroaryl(lower)alkyl,heterocyclyl, substituted heterocyclyl, heteroaryl, or substitutedheteroaryl, each Y is, independently, alkyl, substituted alkyl, cyano,halo, nitro, —SR, —OR, or —NR₂, where each R is, independently,hydrogen, lower alkyl, or substituted lower alkyl, each x is,independently, 0, 1 or 2, and linker connects a carbon designated as cto a carbon designated as d, and is:

 and, where, if R¹ and R² are both —SO₂OH, then: (i) no Y is —SO₂OH;(ii) neither R⁵ nor R⁶ is —SO₂OR⁸ or —OSO₂R⁸; and (iii) R⁵ and R⁶ arenot both selected from the group consisting of hydroxy and hydrogenunless at least one (Y)_(x) is (Y′)_(x′), where x′ is 1 or 2 and Y′ is ahalo, or a pharmaceutically acceptable salt thereof, as a singlestereoisomer or mixture of stereoisomers.
 2. The compound of claim 1, inwhich each x is
 0. 3. The compound of claim 1 that is a compound of theformula:

or a pharmaceutically acceptable salt thereof, as a single stereoisomeror mixture of stereoisomers.
 4. The compound of claim 1, where each Yis, independently, lower alkyl, halo-lower alkyl, lower alkyloxy, cyano,halo, thio, nitro, amino, or hydroxy.
 5. The compound of claim 1, whereR¹ and R² are, independently, —SO₂OR¹⁰, —C(O)OR¹⁰, —SO₂NR¹¹R¹⁰,—C(O)NR¹¹R¹⁰, —OSO₂R¹⁰, —OC(O)R¹⁰, —NR¹¹SO₂R¹⁰, or —NR¹¹C(O)R¹⁰; eachR¹⁰ is, independently, alkyl, substituted alkyl, aryl, substituted aryl,aryl(lower)alkyl, substituted aryl(lower)alkyl, heteroaryl(lower)alkyl,substituted heteroaryl(lower)alkyl, heterocyclyl, substitutedheterocyclyl, heteroaryl, or substituted heteroaryl; and each R¹¹ is,independently, hydrogen or lower alkyl.
 6. The compound of claim 5,where R¹ and R² are, independently, —SO₂NHR¹⁰ or —NHSO₂R¹⁰.
 7. Thecompound of claim 5, where each R¹⁰ is, independently, aryl, heteroaryl,aryl(lower)alkyl, or heteroaryl(lower)alkyl.
 8. The compound of claim 5,where each R¹⁰ is, independently, a substituted alkyl, substituted aryl,substituted aryl(lower)alkyl, substituted heteroaryl(lower)alkyl,substituted heterocyclyl or substituted heteroaryl; at least one of thesubstituents on R¹⁰ is R¹²; each R¹² is, independently, —SO₂OR¹³,—C(O)OR¹³, —SO₂NR¹³ ₂, —C(O)NR¹³ _(2,), hydroxy, triazolyl, tetrazolyl,hydroxyisoxazolyl, a phosphonic acid residue, or a phosphonate residue;and each R¹³ is, independently, hydrogen or lower alkyl.
 9. The compoundof claim 8, where each R¹⁰ is substituted aryl.
 10. The compound ofclaim 9, where each R¹⁰ is substituted phenyl.
 11. The compound of claim10, where each R¹² is, independently, —C(O)OR¹³, —C(O)NR¹³ ₂, hydroxy,triazolyl, tetrazolyl, hydroxyisoxazolyl, a phosphonic acid residue, ora phosphonate residue.
 12. The compound of claim 10, where each R¹² is,independently, —C(O)OR¹³, hydroxy, triazolyl, tetrazolyl,hydroxyisoxazolyl, or a phosphonic acid residue.
 13. The compound ofclaim 10, where each R¹² is, independently, —C(O)OR¹³.
 14. The compoundof claim 13, where R¹² is adjacent on the aryl, heteroaryl, orheterocyclyl ring to a further substituent.
 15. The compound of claim14, where the further substituent is selected from chloro and hydroxy.16. The compound of claim 8, where each R¹² is independently, —SO₂OR¹³or —SO₂NR¹³ ₂.
 17. The compound of claim 16, where each R¹² is,independently, —SO₂OR¹³.
 18. The compound of claim 17, where R¹² isadjacent on the aryl, heteroaryl, or heterocyclyl ring to a furthersubstituent.
 19. The compound of claim 17, where the further substituentis selected from chloro and hydroxy.
 20. The compound of claim 1, whereR¹ is —SO₂OR¹⁰, —C(O)OR¹⁰, —SO₂NR¹¹R¹⁰, —C(O)NR¹¹R₁₀, —OSO₂R¹⁰,—OC(O)R¹⁰, —NR¹¹SO₂R¹⁰, or —NR¹¹C(O)R¹⁰; R² is —SO₂NR¹¹ ₂, —C(O)NR¹¹ ₂,—SO₂OR¹¹, or —C(O)OR¹¹; each R¹⁰ is, independently, alkyl, substitutedalkyl, aryl, substituted aryl, aryl(lower)alkyl, substitutedaryl(lower)alkyl, heteroaryl(lower)alkyl, substitutedheteroaryl(lower)alkyl, heterocyclyl, substituted heterocyclyl,heteroaryl, or substituted heteroaryl; and each R¹¹ is, independently,hydrogen or lower alkyl.
 21. The compound of claim 1, where R¹ and R²are, independently, —C(O)OR⁷ or —C(O)NR⁷ ₂; and each R⁷ is,independently, hydrogen or lower alkyl.
 22. The compound of claim 1,where R¹ and R² are, independently, —SO₂OR⁷or —SO₂NR⁷ ₂; and each R⁷ is,independently, hydrogen or lower alkyl.
 23. The compound of claim 21,where R¹ and R² are —SO₂OH.
 24. The compound of claim 1, where R⁵ and R⁶are, independently, hydrogen, alkyl, substituted alkyl, cyano, halo,nitro, —SR⁸, —OR⁸, or —NR⁸ ₂; and each R⁸ is, independently, hydrogen,lower alkyl, substituted lower alkyl, aryl, substituted aryl,aryl(lower)alkyl, substituted aryl(lower)alkyl, heteroaryl, substitutedheteroaryl, heteroaryl(lower)alkyl, or substitutedheteroaryl(lower)alkyl.
 25. The compound of claim 24, where R⁵ and R⁶are, independently, hydrogen, lower alkyl, halo-lower alkyl, loweralkyloxy, cyano, halo, thio, amino, nitro or hydroxy.
 26. The compoundof claim 1 which is symmetrical.
 27. The compound of claim 1 that isselected from the group consisting of:3-{[(7-{[N-(7-{[(3-sulfophenyl)amino]sulfonyl}-(2-naphthyl))carbamoyl]amino}-2-naphthyl)-sulfonyl]amino}benzenesulfonicacid,3-{[(4-hydroxy-7-{[N-(5-hydroxy-7-{[(3-sulfophenyl)-amino]sulfonyl}-(2-naphthyl))carbamoyl]amino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid,4-{[(7-{[(7-{[(4-sulfophenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)sulfonyl]amino}benzenesulfonicacid, methyl4-({[7-({N-[7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)-2-naphthyl]-carbamoyl}amino)-2-naphthyl]sulfonyl}amino)benzoate,4-{[(7-{[(7-{[(4-carboxyphenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)-sulfonyl]amino}benzoicacid, methyl4-({[4-hydroxy-7-({N-[5-hydroxy-7-({[4-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)-2-naphthyl]sulfonyl}amino)benzoate,4-{[(7-{[(7-{[(4-carboxyphenyl)amino]sulfonyl}-5-hydroxy-(2-naphthyl))amino]carbonylamino}-4-hydroxy-2-naphthyl)sulfonyl]-amino}benzoicacid,3-{[(7-{[N-(7-{[(3-carboxy-2-hydroxyphenyl)amino]sulfonyl}-(2-naphthyl))carbamoyl]-amino}(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoicacid,5-{[(7-{[N-(7-{[(3-carboxy-4-hydroxyphenyl)-amino]sulfonyl}-(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoicacid, methyl3-({[7-({[7-({[3-(methoxycarbonyl)-phenyl]amino}sulfonyl)-2-naphthyl]amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate,3-{[(7-{[(7-{[(3-carboxyphenyl)amino]sulfonyl}-2-naphthyl)amino]carbonylamino}-2-naphthyl)sulfonyl]amino}benzoicacid, methyl3-({[4-hydroxy-7-({[5-hydroxy-7-({[3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)amino}carbonylamino)-2-naphthyl]sulfonyl}amino)benzoate,3-{[(7-{[(7-{[(3-carboxyphenyl)-amino]sulfonyl}-5-hydroxy-(2-naphthyl))amino]carbonylamino}-4-hydroxy-2-naphthyl)-sulfonyl]amino}benzoicacid,5-{[(7-{[N-(7-{[(3-carboxy-4-hydroxyphenyl)-amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}-4-hydroxy(2-naphthyl))sulfonyl]amino}-2-hydroxybenzoicacid,5-{[(7-{[N-(7-{[(3-carboxy-4-chlorophenyl)amino]sulfonyl}-(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]-amino}-2-chlorobenzoicacid,5-{[(7-{[N-(7-{[(3-carboxy-4-chlorophenyl)-amino]sulfonyl}-5-hydroxy(2-naphthyl))carbamoyl]amino}-4-hydroxy(2-naphthyl))sulfonyl]amino}-2-chlorobenzoicacid, methyl2-hydroxy-5-({[7-({[7-({[4-hydroxy-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]amino}carbonylamino)-(2-naphthyl)]sulfonyl}amino)benzoate,methyl2-chloro-5-({[7-({N-[7-({[4-chloro-3-(methoxycarbonyl)phenyl]amino}sulfonyl)(2-naphthyl)]carbamoyl}amino)(2-naphthyl)]sulfonyl}amino)benzoate,N-(7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}-(2-naphthyl))-[(7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))amino]carboxamide,N-(5-hydroxy-7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))[(5-hydroxy-7-{[(3-(1H-1,2,3,4-tetraazol-5-yl)phenyl)amino]sulfonyl}(2-naphthyl))amino]-carboxamide,N-[7-({[3-(diethoxyphosphoryl)-phenyl]amino}sulfonyl)-(2-naphthyl)]{[7-({[3-(diethoxyphosphoryl)phenyl]amino}sulfonyl)-(2-naphthyl)]amino}carboxamide,N-[7-({[3-(ethoxy(hydroxyphosphoryl))-phenyl]amino}sulfonyl)-(2-naphthyl)]-{[7-({[3-(ethoxy(hydroxyphosphoryl))-phenyl]amino}sulfonyl)(2-naphthyl)]amino}carboxamide,2-chloro-5-{[(7-{[(7-{[(4-chloro-3-sulfophenyl)amino]sulfonyl}-2-naphthyl))amino]-carbonylamino}(2-naphthyl))sulfonyl]amino}-benzenesulfonicacid,2-chloro-5-{[(7-{[(7-{[(4-chloro-3-sulfophenyl)amino]sulfonyl}-5-hydroxy(2-naphthyl))amino]-carbonylamino}-4-hydroxy-(2-naphthyl))sulfonyl]amino}benzenesulfonicacid,5-[({7-[3-(7-{[(3-carboxy-4-hydroxyphenyl)amino]sulfonyl}-(2-naphthyl))-2-oxoimidazolidinyl](2-naphthyl)}sulfonyl)amino]-2-hydroxybenzoicacid,5-[({7-[3-(7-{[(3-carboxy-4-chlorophenyl)-amino]sulfonyl}-(2-naphthyl))-2-oxoimidazolidinyl](2-naphthyl)}-sulfonyl)amino]-2-chlorobenzoicacid,2-chloro-5-{[(7-{[N-(5-hydroxy-7-{[(3-sulfophenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]-amino}benzoicacid,5-{[(7-{[N-(7-{[(4-carboxyphenyl)amino]sulfonyl}(2-naphthyl))carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-chlorobenzoicacid;5-{[(7-{[N-(7-{[(3-carboxyphenyl)amino]sulfonyl}(2-naphthyl))-carbamoyl]amino}(2-naphthyl))sulfonyl]amino}-2-chlorobenzoicacid, and the pharmaceutically acceptable salts thereof.
 28. Apharmaceutical composition for treating a mammalian disease stateselected from the group consisting of hyperglycemia, type I diabetes,and type II diabetes, comprising: (a) a therapeutically effective amountof a compound of claim 1; and (b) at least one pharmaceuticallyacceptable excipient.
 29. A method of stimulating the kinase activity ofthe insulin receptor, comprising contacting the insulin receptor, or thekinase portion thereof, with a compound of claim 1 in an amountsufficient to stimulate the kinase activity of the insulin receptor. 30.A method of activating the insulin receptor, comprising contacting theinsulin receptor, or the kinase portion thereof, with a compound ofclaim 1, in an amount sufficient to activate the insulin receptor.
 31. Amethod of stimulating the uptake of glucose into cells displaying theinsulin receptor, comprising contacting the cells with a compound ofclaim 1 in an amount sufficient to stimulate the uptake of glucose intothe cells.
 32. A method of treating a disease state in a mammal selectedfrom the group consisting of hyperglycemia, type I diabetes, and type IIdiabetes, comprising administering a therapeutically effective amount ofa compound of claim 1, or a pharmaceutical composition thereof, to themammal.
 33. The method of claim 32, further comprising treating saidmammal with an additional form of therapy for said disease state. 34.The radiolabeled compound of claim
 1. 35. The compound of claim 34 thatis¹⁴C-4-methylphenyl-3-[({4-hydroxy-7-[(N-{5-hydroxy-7-[({3-[(4-methylphenyl)oxysulfonyl]phenyl}amino)sulfonyl](2-naphthyl)}carbamoyl)amino]2-naphthyl}sulfonyl)amino]benzenesulfonate.36. A method of identifying a compound that has the function ofstimulating the kinase activity of the insulin receptor, activating theinsulin receptor, and stimulating the uptake of glucose, comprisingusing the compound of claim 35 as a diagnostic.
 37. A process forpreparing a compound which mimics the function of the compound of claim1, comprising: (i) submitting a test compound to a screen fordetermining its stimulation of the kinase activity of the insulinreceptor in relation to a compound of claim 1; and (ii) preparing thetest compound if it exhibits stimulation of the kinase activity of theinsulin receptor.